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. 2005 Oct 10;171(1):61–73. doi: 10.1083/jcb.200502078

Figure 1.

Figure 1.

Activation of Mkk3/p38 MAPK is necessary for CHOP induction in FC-loaded macrophages. (A) WT and Mkk3 − / − macrophages (Mφs) were FC-loaded for 0, 2, 5, 7, 10, and 15 h using 100 μg/ml ac-LDL plus the ACAT inhibitor 58035. Whole cell lysates were prepared as described under “Materials and methods” and immunoblotted for CHOP (top panel), activated phospho-Thr180/Tyr182 p38 MAPK (phospho-p38, middle panel), and total p38 (bottom panel). Lines indicate areas of the same gel that were spliced together. (B) WT and Chop − / − macrophages were left untreated (−) or were FC-loaded (+) for 5 h, and lysates were immunoblotted for phospho-p38 and total p38. (C) WT and Mkk3 − / − macrophages were left untreated (−) or were FC-loaded for 7 h (+). In some experiments, WT cells were treated with ac-LDL alone, which showed only minimal MK2 phosphorylation (not depicted). Whole cell lysates were immunoblotted for activated phospho-Thr334 MK2 (phospho-MK2, top panel) and total MK2 (bottom panel). (D) WT and Mk2 − / − macrophages were left untreated (−) or were FC-loaded for 7 h (+). Whole cell lysates were immunoblotted for CHOP (top panel), total MK2 (second panel), total p38 (third panel), or tubulin as a loading control (bottom panel). Lines indicate areas of the same gel that were spliced together.