Figure 4.

Statins did not change cell viability, or expression of PKC isozymes or their functions. (a) Statin treatment did not change cell viability. Left, representative images of HEK 293 T cells treated overnight with 3 μM of statins as indicated. Cells were labeled after staining with Trypan blue for viability count. Control cells were treated with either 0.01% DMSO (CTRL) or 8% ethanol. Right, summary data of experiments performed to the left. (b) Statin treatment did not change cPKC expression level. Typical immuno-blot and summary data of endogenous cPKC expression in HEK293T cells after 2 µM statin treatment for overnight. cPKC isozymes are recognized by an anti-cPKC antibody. *p = 0.05 (c) Typical confocal images of HEK293T cells expressing GFP tagged PKCα before and after acute phenylephrine treatment (within 1 minute) for stimulation of overexpressing α1A-AR (Phe, 30 µM), in the presence and absence of overnight fluvastatin treatment (2 µM) as indicated. Fluorescence profile was measured at the cell cross section indicated by the yellow line and shown in the inset to the right. M and C indicate plasma membrane and cytoplasm respectively. (d) Top, typical confocal images of PKCα-GFP, PKCβI-GFP, PKCβII-GFP and PKCε-GFP before and after acute Phe treatment (30 μM, details in Methods) at the absence and presence of overnight statins treatment (all at 2 μM). Bottom, summary of the translocation of PKC isozymes in experiments conducted as above. *p < 0.05 (number of cells analyzed).