Computational protocol: PPM1A silences cytosolic RNA sensing and antiviral defense through direct dephosphorylation of MAVS and TBK1

Similar protocols

Protocol publication

[…] To visualize the subcellular localization of TBK1, IRF3, and PPM1A, HaCaT or HeLa cells were treated as indicated in the figures, fixed in paraformaldehyde, permeabilized, and blocked by horse serum. Cell sides were then incubated sequentially with primary antibodies (anti-IRF3, anti-PPM1A, or anti-Myc) and Alexa-labeled secondary antibodies, followed by extensive washing. Slides were then mounted with VECTASHIELD and stained with DAPI (Vector Laboratories). Mitochondria were visualized by MitoSox Red (Life). Immunofluorescence images were obtained and analyzed using a Nikon Eclipse Ti inverted microscope, or GE DeltaVision OMX system, and ImageJ software. [...] Quantitative data are presented as means ± SEM from at least three independent experiments. Data presented as either fold change or percentage were log-transformed before statistical analysis. When appropriate, statistical differences between groups were analyzed using an unpaired or paired Student’s t test or by SigmaPlot 10.0. Differences were considered significant at P < 0.05. […]

Pipeline specifications

Software tools ImageJ, SigmaPlot
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Danio rerio, Mus musculus, Homo sapiens
Chemicals Tretinoin