miR-155 Regulated Inflammation Response by the SOCS1-STAT3-PDCD4 Axis in Atherogenesis

Figure 6

SOCS1-STAT3 pathway regulated PDCD4. (a) Fluorescence images (×100) showed RAW264.7 cells were infected with SOCS1 overexpression adenovirus (Adv. SOCS1) and vector adenovirus (Adv. GFP). (b) qPCR detected the SOCS1 mRNA expression after cells were infected with Adv. SOCS1 and Adv. GFP for 48 h: versus control, versus Adv. GFP group. (c) Downregulation of the expression of STAT3 mRNA by STAT3 siRNA in macrophages RAW264.7 cells. The relative expression of STAT3 mRNA was detected by qPCR after 50pM STAT3 siRNA, including si-01, si-02, and si-03, and negative control (NC) was transfected into RAW264.7 cells for 48 h, respectively: versus control, versus NC group. (d) Western blot analyzed the protein level of phospho-STAT3 (p-STAT3) when RAW264.7 cells had been treated by 50pM STAT3 siRNA (si-03) and NC for 48 h. Representative bands show the protein expression of STAT3 (the upper bands), β-actin (the lower bands) in left panel, and the histograms in right panel showing quantification of STAT3 bands’ normalization relative to β-actin expression in (d). versus NC group. (e) Western blot analyzed the protein level of SOCS1, p-STAT3, and PDCD4 in RAW264.7 cells. They were transfected into the Adv. SOCS1 or STAT3 siRNA for 24 h and then treated by ox-LDL for 24 h. Representative bands show the protein expression of SOCS1 (the upper bands), p-STAT3 (the middle bands), PDCD4 (the third bands), and β-actin (the lower bands) in left panel, and the histograms in right panel showing quantification of STAT3 bands’ normalization relative to β-actin expression. versus control, versus 20 μg/mL ox-LDL group, and versus 20 μg/mL ox-LDL plus Adv. SOCS1 group. The data are presented as the mean ± SE of three separate experiments.

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