Figure 6: CHIP degrades SHP-1 through ubiquitin-proteasome system and activates STAT3 and ERK1/2 signaling pathway. | Scientific Reports

Figure 6: CHIP degrades SHP-1 through ubiquitin-proteasome system and activates STAT3 and ERK1/2 signaling pathway.

From: Protection against doxorubicin-induced myocardial dysfunction in mice by cardiac-specific expression of carboxyl terminus of hsp70-interacting protein

Figure 6

(a) Western blot analysis of protein levels of p53, SHP-1, p-STAT3/STAT3 and p-ERK1/2/ERK1/2 in WT and CHIP-TG hearts (left). Quantitative analysis of relative protein levels (right). *P < 0.05, **P < 0.01 versus WT mice + saline; #P < 0.05 versus WT mice + DOX. (b) The interactions of SHP-1 with CHIP were detected with co-immunoprecipitation in H9c2 cells. H9c2 cells lysates were immune-precipitated with anti-CHIP antibody or control IgG, and the immune-precipitates were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotted with anti-SHP-1 antibody. (c) Equal amounts of lysates from Ad-GFP/Ad-CHIP adenovirus H9c2 cells were immune-precipitated with SHP-1 antibody and analyzed by immunoblotted with anti-ubiquitin antibody to detect ubiquitylated forms of SHP-1 in vitro. (d) Western blot analysis of protein levels of SHP-1, p-STAT3/STAT3 and p-ERK1/2/ERK1/2 in H9c2 cells after transfected with GFP/CHIP adenovirus and treated with or without MG 132 (left). Quantitative analysis of relative protein levels (right). (e) Western blot analysis of protein levels of SHP-1, p-Jak/Jak, p-STAT3/STAT3 and p-ERK1/2/ERK1/2 in H9c2 cells after transfected with siRNA-control, siRNA-CHIP or siRNA-SHP-1 (left). Quantitative analysis of relative protein levels (right). *P < 0.05, **P < 0.01, ***P < 0.001 versus Ad-GFP/siRNA-control; #P < 0.05, ###P < 0.001 versus Ad-GFP + MG132/siRNA-CHIP.

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