Skip to main content
. Author manuscript; available in PMC: 2012 Aug 9.
Published in final edited form as: Cell Signal. 2010 Mar 7;22(7):1063–1075. doi: 10.1016/j.cellsig.2010.02.009

Figure 4. Activation of p38 MAPK induces translocation of endogenous MK5 in HEK293 cells.

Figure 4

HEK293 cells were transfected with pIRES-MK5.1-V5-EGFP. After 24 h cells were serum starved for 6 h and then treated with or without sorbitol (0.3 M) (A) or anisomycin (15 μg/ml) (B) for the indicated times and then lysates were prepared. Activation of p38 was detected by immunoblotting using an anti-phospho p38 antibody. The stability of exogenous MK5 and endogenous p38α during prolonged stress was verified by immunoblotting using anti-V5 and anti-p38α antibodies. (C) Untransfected HEK293 cells were grown for 48 hours, serum starved for 6 h, and then treated with or without sorbitol (0.3 M) or anisomycin (15 μg/ml) for 2 h. Cells were then fixed and MK5 was visualized by staining with an anti-MK5 antibody and an Alexa 555-coupled secondary (anti-mouse) antibody. The upper panels are the fluorescence images whereas the lower panels contain differential interference contrast images of the same cells. Images were acquired using a 63x/1.4 plan-apochromat oil DIC objective lens.