FIGURE 5:

CCTα−S315D+S319D is partially defective in localization to nuclear membranes. (A) CHO-MT58 cells transiently expressing wild-type CCTα and the indicated CCTα phosphomutants were cultured at 40°C and treated with 300 µM oleate/BSA for 60 min, fixed, and immunostained with CCTα and LMNA/C primary antibodies followed by AlexaFluor–488 and –594 secondary antibodies, respectively. All images are 0.8-μm confocal sections; bar, 10 μm. (B) Lysates prepared from CHO-MT58 treated as described in A were immunoblotted with CCTα, CCTα/β-pS319, CCTα-pY359+pS362, and actin antibodies. Confocal images and immunoblot are representative of three experiments.