Rotavirus cell infection and immunofluorescence. MA-104 Clone 1 cells (American Type Culture Collection; ATCC:CRL-2378.1; RRID:CVCL_3846) were cultured in DMEM-RS media supplemented with 5% fetal bovine serum at 37°C and 5% CO2. Prior to infection, Rhesus rotavirus (RRV) was activated with trypsin (10 μg/ml) for 30 min at 37°C. MA104 cells grown on glass coverslips were infected with RRV at a multiplicity of infection (MOI) of 1. Cells were fixed and prepared for immunofluorescence after six hours post-infection. Using a STORM imaging buffer mounting medium (1.5% glucose oxidase + 100 mM β-mercaptoethanol), the coverslips were mounted onto the center of glass slides. Rotavirus replication machinery imaging. Cell infection micrographs were provided kindly by Garcés and collaborators [1]. Imaging was carried out with an Olympus IX-81 inverted microscope in TIRF mode (Olympus, cellTIRFM illuminator), with an evanescence field depth of 200 nm. The objective lens used was an Olympus UApo N 100x 1.4 NA oil-immersion, with an additional 1.6x magnification lens. Excitation of Alexa-488 and Alexa-568 was provided by 488 nm and 568 nm lasers, respectively. Using a laser-modulation protocol, previously described in [1], an EMCCD camera (iXon 897, Model No: DU-897E-CS0-#BV; Andor) was used to acquire the images. Acquisition rate was set to 20 fps with a pixel size of 100 nm. For processing with MSSR, the algorithm parameters were set as follows: AMP = 5, PSF = 3, Order = 1, GPU parallel computing = enabled, temporal analysis = Mean (100 frames). 1. Garcés S, Y. et al. Nanoscale organization of rotavirus replication machineries. elife 8, e42906, (2019).

For all files, the red channel corresponds to the NSP2 protein, while the green channel corresponds to the viral protein depicted by each file name.

{"references": ["Garc\u00e9s S, Y. et al. Nanoscale organization of rotavirus replication machineries. elife 8, e42906, (2019)."]}