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. 2021 Dec 30;14(1):60. doi: 10.3390/v14010060

Figure 6.

Figure 6

GPS491 alters expression and phosphorylation of selected SR proteins. HeLa rtTA HIV∆mls cells were treated with DMSO or GPS491 (2.5 µM) and induced with Dox. Twenty hours post induction, cells were harvested in RIPA buffer containing a phosphatase inhibitor cocktail and fractionated on 10% stain-free SDS-PAGE gels. (a) Representative Western blots for each SR protein are shown. At the right, a summary of Western blots from three independent assays measuring the effect of GPS491 on SR proteins levels relative to DMSO-treated cells. Asterisks (*) represent changes in expression level at p < 0.05. (b) Cell lysates were mock-treated or treated with alkaline phosphatase (PPase) prior to loading onto 10% stain-free gels. After transfer onto PVDF, blots were probed for SRSF4. Blots are representative of n > 3 independent assays. (c) Cells treated with DMSO or GPS491 were fixed and permeabilized followed by staining with antibodies to either SRSF2 or SRSF4. DAPI was used to stain for cell nuclei. Images were captured on Leica DMR Microscope at 63× oil immersion.