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Fig. 2 | Cell & Bioscience

Fig. 2

From: Hypoxia induces calpain activity and degrades SMAD2 to attenuate TGFβ signaling in macrophages

Fig. 2

Expression of TGFBR2 and SMAD7 in response to TGFß/hypoxia. Macrophages were transfected with non-targeting siRNA constructs (ctr), siRNA-HIF1α (si1α), or siRNA-HIF2α (si2α) and mRNA expression of TGFBR2 a or SMAD7 b were analyzed by quantitative PCR after treatments with TGFß under normoxia (nor) or hypoxia (hy) for 8 h. c Cell surface expression of TGFBR2 was followed by flow cytometry after stimulation of macrophages with TGFß under normoxia (nor) vs. hypoxia (hy) for 8 h and d mean fluorescence intensity was calculated from 4 individual experiments. e, f Time-dependent Western blot analysis and statistical evaluation of SMAD7 expression in macrophages exposed to TGFß under normoxia or hypoxia. g, h Macrophages were exposed to the phosphatase inhibitor okadaic acid (OA) 1 h prior to adding TGFß for 8 h. Phosphorylation and total protein expression of SMAD2 was followed by Western blot analysis, using tubulin as a loading control

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