IFGII Induced Overexpression of H-Ras in the Mesenchymal Stem Cells
(A) Anti-IGFII co- IP followed by western blotting with anti-P62,anti-TNFR1, anti-CLYD, anti-EGR1, anti-NFκB, anti-TLR4, and anti-PPARγ in the TNF-α-treated mesenchymal stem cells transfected with pCMV6-AC-GFP, pCMV6-AC-GFP-P62, pGFP-V-RS, pGFP-V-RS-P62, and IgG IP as negative control. INPUT refers to western blotting with anti-IGFII. (B) CHIP with anti-IGFII, anti-P62, anti-TNFR1, anti-CLYD, anti-NFκB, or anti-PPARγ followed by PCR with H-Ras promoter primer in the TNF-α-treated mesenchymal stem cells transfected with pCMV6-AC-GFP (GFP-control), pCMV6-AC-GFP-P62 (GFP-P62), pGFP-V-RS (RNAi control), and pGFP-V-RS-P62 (P62 RNAi). IgG CHIP was used as negative control. H-Ras promoter as INPUT is shown. (C) The H-Ras promoter luciferase activity assay in the TNF-α-treated mesenchymal stem cells transfected with pCMV6-AC-GFP (GFP-control), pCMV6-AC-GFP-P62 (GFP-P62), pGFP-V-RS (RNAi control), or pGFP-V-RS-P62 (P62 RNAi). (D) RT-PCR with H-Ras primers in the TNF-α-treated mesenchymal stem cells transfected with pCMV6-AC-GFP (GFP-control), pCMV6-AC-GFP-P62 (GFP-P62), pGFP-V-RS (RNAi control), and pGFP-V-RS-P62 (P62 RNAi), respectively. β-actin as internal control is shown. (E) Western blotting with anti-H-Ras and anti-pH-Ras in the TNF-α-treated mesenchymal stem cells transfected with pCMV6-AC-GFP (GFP-control), pCMV6- AC-GFP-P62 (GFP-P62), pGFP-V-RS (RNAi control), and pGFP-V-RS-P62 (P62 RNAi), respectively. β-actin was used as an internal control. **p < 0.01.