Additional file 2. Supplemental Table 2: DEPICT genetic association in pathways.
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Additional file 6. Autocorrelation z scores between pairs of stimulated and unstimulated chromatograms.
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Spreadsheet of analyte concentrations and metabolite/parent ratios for tamoxifen and its metabolites. (XLSX 10 kb)
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Additional file 5: Table S4. EGFR information on LGSC cell lines.
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Article full textThe above summary slide represents the opinions of the authors. For a full list of declarations, including funding and author disclosure statements, please see the full text online (see “read the peer-reviewed publication” opposite). © The authors, CC-BY-NC 2021.
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Table S1. Next-generation sequencing results of erythrocyte microRNAs for 9 healthy controls and 9 relapsing-remitting Multiple Sclerosis patients. (XLSX 27Â kb)
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Background: The use of cannabis is highly prevalent during adolescence compared to the general adult population. In addition to the high comorbidity between cannabis use and anxiety disorders, early evidence suggests that cannabis may precede the development of anxiety. Moreover, adolescence represents a major developmental period for both neurobiological and psychological processes, placing these individuals at a heightened vulnerability to the influence of cannabis. Objectives: This systematic review and meta-analysis examined the prospective associations between adolescent cannabis use and subsequent anxiety outcomes (i.e. anxiety disorders and/or symptoms). Methods: Following PRISMA guidelines, a systematic review and meta-analysis were conducted encompassing data from articles published between database inception and September 2022. Results: Six longitudinal studies were identified for quantitative analysis, while twelve non-overlapping longitudinal studies were identified for qualitative review (total N = 18; 33380 subjects). Meta-analytical findings supported an association between adolescent cannabis use and the development of a subsequent anxiety disorder (Odds Ratio = 2.14, 95% CI: 1.37–3.36, p < .01). These findings were consistent with our qualitative synthesis where nine of the twelve longitudinal studies observed a significant relationship between adolescent cannabis use and exacerbation of anxiety symptoms later in life, irrespective of an anxiety disorder diagnosis. Discussion: In summary, the current evidence suggests a prospective association between adolescent cannabis use and later anxiety symptoms and disorders. These findings underscore the importance of refining research methodologies, considering sex-based differences and controlling for confounding factors, as well as implementing educational initiatives and developing clinical interventions to address the mental health risks associated with cannabis use among adolescents.
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Extended Data Fig. 1 | Predicted PLD in Pex13/Pex5. PLD Analysis of a, Pex13 b, Pex5 and c, Pex14 using PLAAC21. Predicted PLD sequence is marked in red for Pex13 and Pex5 on the left, corresponding to the outputs on the right. d, Multiple sequence alignment of the Pex13 sequence from the indicated five species, showing conserved Y residues within the predicted yeast PLD sequence. Extended Fig. 2. PTS1 transport defective in Pex13-PLD variants a, Distribution of Y (white) to F (orange) or S (purple) substitutions introduced in the Pex13 PLD using CRISPR-Cas9. Point mutation heat map shown in shades of gray denotes the number of mCherry foci observed in the indicated cells. n = 100 cells from 3 independent experiments. Confocal images of b, GFP-Pex13-S8 (interspersed) and c, GFP-Pex13-S9 (blocky) cells taken at the indicated time points after induction of Pex5. Scale bar 5 mm. Percentage cells displaying mCherry-SKL foci are indicated below each panel. n = 100 cells per sample. Extended Data Fig. 3 | Import kinetics in Pex13-WT and Y→S/F mutants. a, Percentage of cells displaying mCherry-SKL foci in the indicated strains following Pex5 induction. Error bars are mean ± s.e.m. for 3 independent experiments, n=100 cells. The schematic on the right shows the Y→S/F substitutions made. b, Timelapse following mCherry-SKL foci formation in Pex13 WT cells (black arrow) and c, Pex13 S8 cells (orange/blue arrows). Scale bar 5 µm. d, e, mCherry-SKL foci fluorescence intensity of the cells imaged in panels b and c, respectively, plotted as a function of time after induction of Pex5 expression. Extended Data Fig. 4 | Representative images of cargo import defects. Spinning disc confocal images of a, GFP-Pex13-S7 and b, GFP-Pex13-S15 cells taken at the indicated time points after induction of Pex5. Scale bar 5 µm. Percentage cells displaying mCherry-SKL foci are indicated below the panels. n = 100 cells. Extended Data Fig. 5 | LLPS of purified PTS1 peroxins. a, Coomassie stained SDS-PAGE gels of 1 µg of the indicated purified proteins. Images of condensates formed by b,c, Pex13-IDR WT labeled with AZdye 488 maleimide d,e, Pex5 labeled with AZdye 647 maleimide and f,g, Pex14 IDR labeled with AZdye 488 maleimide. Scale bar 5 µm. Extended Data Fig. 6 | Pex5-cargo partitions into Pex13-IDR condensates. Titration of a, Pex5 and b, mCherry-SKL to determine the concentration at which they do not form condensates. 1 µM of both Pex5 and mCherry-SKL were used for in vitro reconstitution assays with Pex13-IDR. c, Reconstitution experiments to measure partitioning of Pex5 labelled with AZ 647 dye and mCherry-SKL into Pex13-IDR WT condensates immediately after mixing at room temperature. d, Cas9-AZdye 488 maleimide does not actively partition into Pex13 droplets (top) while mCherry-SKL partitions with Pex5 and Pex13 (bottom). e, Pex5 and mCherry-SKL partition into Pex13-IDR WT (10 µM) and Pex13-IDR S8 (30 µM) condensates. f, Pex13-IDR S15 does not form spherical condensates. mCherry-SKL remains diffuse in most images of Pex13-S15 aggregates (top). In some images, however, Pex5-mCherry-SKL appears to partition with Pex13-S15 (bottom). This is likely due to experimental variation, perhaps a somewhat lower concentration of Pex5-mCherry-SKL, co-aggregating with Pex13-IDR-S15. Scale bar 5 µm. Extended Data Fig. 7 | iFCCS) workflow and controls a, Fiducial markers used to align the GFP-Pex13 and mCherry-SKL channels using MATLAB b, The centroid position of fiducial markers are selected and the coordinates are exported using a normalized cross correlation between the red and green channels. c, The centroid locations of peroxisomes of interest are localized using a particle localization analysis (dashed circle provided as guide for the eye). d, The GFP and mCherry intensities of each of the surrounding 17x17 pixels of the peroxisome centroid location are extracted. The intensities of centroid pixel boxed in c are shown in d. Using the equation in e, the intensity fluctuations, δF(t), relative to the mean intensity are calculated and used to calculate G(τ). Resulting data f, Spatial and g, Temporal, as explained in the methods section. h, Controls for laser power effect on photo bleaching. i, Averaged standard deviation of the cross correlation curves with the maximum G_XC(0) value in each peroxisome at each laser power. Extended Data Fig. 8 | iFCCS spatial and temporal data shows transient clusters of correlated signal of mCherry-SKL and GFP-Pex13. a, Percentage of peroxisomes with GXC(0)>0.5 for GFP-Pex13 and soluble mCherry (no SKL) (gray). The-data of GFP-Pex13 with mCherry-SKL import from Fig. 4j is shown for comparison (red). b, Spatial iFCCS data from six different peroxisomes in WT 50 min after Pex5 induction where distinct transient clusters of correlated signals are observed. Images measure 1.19 µm x 1.19 µm in size. c, Transient nature of individual clusters are observed in the decay and fluctuations of the cross-correlation over time between clusters in different peroxisomes (n=27 peroxisomes). d, Gaussian distribution of NGFP-Pex13/N-mCherry-SKL obtained by autocorrelating signal from the Pex13-GFP and SKL-mCherry channels, extracting a value from the pixel exhibiting the highest GXC(0) > 0.5 in each peroxisome from the Pex13 WT strain, 50 min after Pex5 induction. e, Bimodal Gaussian fit of GFP-Pex14 same as in panel d, for data obtained 90 min after Pex5 induction. f, Distribution of NPex13-488/NPex5-647 showing the ratio of Pex13 to Pex5 in Pex13-IDR and Pex13-S8 condensates. All pixels within a single condensate of Pex13-IDR-WT or Pex13-IDR-S8 with Pex5 partitioned within the condensate were analyzed. Gaussian fit parameters µ and σ listed with 95% confidence intervals in panels d,e and f. g, An alternative transmembrane intercalation model for peroxisomal cargo import.
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Table S1A. All hydrogen deuterium exchange (HDX) peptide data for experiments examining the global exchange of PI4KIIIA, TTC7B, and FAM126A. The charge state (Z), residue start, residue end number, retention time (RT) and sequence are displayed for every peptide. In the Raw Data column, the two time points (0.3s and full) are labelled, and the relative level of HDX is coloured according to the amount of deuterium incorporated, on a blue to red continuum. The data listed for the 0.3s time point are the average of three independent experiments, with SD shown next to all HDX values. In the Normalized to Full Deuteration column, the 0.3s data has been normalised to the full deuteration measurements with the exception of those data (surrounded by black lines) where the full deuteration measurement was lower than 20% deuterium incorporation. The third column denotes the corresponding peptide centroid. Table S1B. All HDX peptide data for experiments examining the complex dynamics of PI4KIIIA, TTC7B, and FAM126A. The charge state (Z), residue start, residue end number, retention time (RT) and sequence are displayed for every peptide. The two columns represent each state examined (+/- PI4KIIIA) and contain the data for five time points. The data listed are the average of three independent experiments, with SD shown next to all HDX values. Table S1C. All HDX peptide data for experiments examining the dynamics of inhibitor specificity of PI4KIIIA, TTC7B, and FAM126A. The charge state (Z), residue start, residue end number, retention time (RT) and sequence are displayed for every peptide. The three columns represent each state examined (+/- inhibitor) and contain the data for four time points. The data listed are the average of three independent experiments, with SD shown next to all HDX values.
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This dataset contains the raw, unprocessed immunofluorescence, live-cell imaging and Western blot data for the manuscript entitled: Interactome rewiring following pharmacological targeting of BET bromodomains. All methods employed to generate this dataset can be found in the accompanying manuscript and supplemental material.
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Additional file 2. Supplemental Table 2: DEPICT genetic association in pathways.
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Additional file 6. Autocorrelation z scores between pairs of stimulated and unstimulated chromatograms.
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Spreadsheet of analyte concentrations and metabolite/parent ratios for tamoxifen and its metabolites. (XLSX 10 kb)
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