Figure 2.

Localization of p38 in dot-like structures is specific for the interaction between Gadd45 proteins and MEKK4. (a) NIH/3T3 cells were transiently transfected with either V5-tagged Gadd45α, V5-tagged Gadd45β or V5-tagged Gadd45γ together with GFP-tagged LC3 and HA-tagged MEKK4. At 24 h post transfection, the cells were stained with anti-V5 (blue), LC3-GFP (green) and anti-phospho-p38 (red). Subsequently, samples were analyzed by confocal microscopy. Bars, 10 μm. (b) NIH/3T3 cells were transiently transfected with V5-tagged Gadd45β and HA-tagged ASK1. At 24 h after transfection, the staining was performed with anti-V5 (green), anti-HA (red) and endogenous phosphorylated p38 (blue). Subsequently, samples were analyzed by confocal microscopy. Bars, 10 μm. (c) NIH/3T3 cells were transfected with V5-tagged Gadd45β HA-tagged ASK1 and GFP-LC3. After 18 h, cells were fixed and stained with anti-HA (red) and anti-V5 antibody (blue). Bars, 10 μm