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ZENODO
Dataset . 2024
License: CC BY
Data sources: ZENODO
ZENODO
Dataset . 2024
License: CC BY
Data sources: Datacite
ZENODO
Dataset . 2024
License: CC BY
Data sources: Datacite
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Developing nanobodies as allosteric molecular chaperones of glucocerebrosidase function

Authors: Dal Maso, Thomas; Plotegher, Nicoletta; Versées, Wim;

Developing nanobodies as allosteric molecular chaperones of glucocerebrosidase function

Abstract

OVERVIEW OF THE RAW DATA FILES - DalMaso_et_al_Fig1A.xlsx: Raw data of the ELISA of 20 purified Nbs using GCase or N370S GCase coated on the bottom of the ELISA wells. - DalMaso_et_al_Fig1C.xlsx: KD table of Nbs for GCase and N370S GCase. - DalMaso_et_al_Fig 2B 2C: Results of GCase activity assay performed with 20 Nbs incubated with either velaglucerase or cellular lysates. - DalMaso_et_al_Fig2E.xlsx: Thermal unfolding curves of GCase, obtained using a thermal shift assay, in absence or presence of Nbs. - DalMaso_et_al_Fig4B 4C: Cytograms and raw data of GCase activity in live HEK293T cells transfected with ER- and lysosomal-targeted- Nbs. - DalMaso_et_al_Fig4E 4F: Cytograms and raw data of lysosomal proteolytic activity evaluated in HEK293T cells transfected with ER- and lysosomal-targeted Nbs by DQBSA assay. - DalMaso_et_al_Fig5A 5B: Western blotting of lysate of Nb-transfected HEK293T cells incubated with ENDO H and PNGase F. - DalMaso_et_al_Fig5C 5D: Confocal images of HEK293T cells transfected with Nb1/Nb4/Nb9- flag-IRES-EGFP trageted to the ER stained with flag and calnexin for colocazation Nb-flag/ER-CALNEXIN. Pearson's coefficients calculated using Jacop plugin of ImageJ between FLAG and CALNEXIN. - DalMaso_et_al_Fig5e 5f: Confocal images of HEK293T cells transfected with Nb1/Nb4/Nb9- flag-IRES-EGFP trageted to the ER stained with flag and LAMP2A for colocazation Nb-flag/lysosomes LAMP2A. Pearson's coefficients calculated using Jacop plugin of ImageJ between FLAG and LAMP2A. - DalMaso_et_al_Fig6A 6B S15: Raw data of 4-MU assay performed with 20 Nbs incubated either with recombinant N370S or with tissue lysates from Gba-/- hN370S mice. Results of 4-MU assay perfromed on gut lysates from wild-type, heterozygous and knock-out mice. - DalMaso_et_al_Fig6C 6D S16E: Cytograms and raw data of GCase activity evaluated in live HEK GBA1 knockdown cells co-transfected with N370S GCase mutant and ER-Nbs and Lysosomal-Nbs. Cytograms of GCase activity in live wild-type, 11F GBA1 knock-down cell clone and 11F GBA1 knock-down overexpressing N370S mutant. - DalMaso_et_al_FigS3.xlsx: Raw data of the ELISA of 20 purified Nbs using GCase or deglycosylated GCase coated on the bottom of the ELISA wells. - DalMaso_et_al_FigS4.xlsx: Biolayer interferometry (BLI) sensorgrams data for the titration of GCase into a set of Nbs. - DalMaso_et_al_FigS5.xlsx: Fluorescence polarisation raw data for Nb16-5TAMRA and GCase. - DalMaso_et_al_FigS6.xlsx: Biolayer interferometry (BLI) sensorgrams data of GCase binding to immobilized Nbs (reverse set-up). - DalMaso_et_al_FigS7.xlsx: Cathepsin L proteolysis assay raw data (western blot and quantification). - DalMaso_et_al_FigS14.xlsx: Biolayer interferometry (BLI) sensorgrams data for the titration of GCase N370S into a set of Nbs. - DalMaso_et_al_FigS10A S10B S11A S11B: western blotting of HEK293T cells transfected with Mock and ER- and lysosomal targeted Nbs to evaluate Nbs expression (FLAG) and their impact on ER (CALNEXIN) or lysosomal (LAMP1) compartments and on GCase expression (GCase) - DalMaso_et_al_FigS10E: confocal images HEK293T cells overexpressing Mock sequence, Nb1, Nb4 and Nb9 stained with an anti-flag antibody, either targeted to the ER or to the lysosomes, with the relative EGFP fluorescence signal. - DalMaso_et_al_FigS12A: cytograms and raw data of GCase activity in live HEK transfected with ER- and lysosomal Nbs and treated with GCase inhibitor CBE to evaluate the non-specific signal due to PFB-FDGlu off-target hydrolysis, or to other non-specific effects. - DalMaso_et_al_Figs13: uncropped SDS-PAGE and blot of N370S GCase purified from 293T Freestyle cells - DalMaso_et_al_FigS16A S16B: western blotting of 6E and 11F GBA1 knock-down cell clones to evaluate GCase expression. - DalMaso_et_al_FigS16C: cytograms of GCase activity in live wild-type and 6E and 11F GBA1 knock-down cell clones - DalMaso_et_al_FigS16D: confocal images of 11F GBA1 knock down cell clone co-transfected with N370S GCase mutant and ER-targeted Nb4 (choosed as representative).

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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
0
Average
Average
Average