Figure 3.

EGR1 mediated the promoting fibrosis role of MBD2 during BUMPT cells treated with TGF-β1 or co-culture of murine embryonic NIH 3T3 fibroblasts, and BUMPT cells

BUMPT cells were treated with 5 ng/mL TGF-β1 for 0–48 h or transfected with EGR1 siRNA and then treated with 5 ng/mL TGF-β1 for 24 h. (A) Immunoblot analysis of EGR1 at indicated time points. (B) Analysis of the gray-scale image between them. (C) Immunoblot analysis of EGR1, FN, and Col I and IV. (D) Analysis of the gray-scale image between them. (E) Immunoblot analysis of SMAD3, p-SMAD3, AP-1, TGF-β, ERK1/2, and p-ERK1/2. (F) Analysis of the gray-scale image between them. (G) The murine embryonic NIH 3T3 fibroblasts were treated with the supernatant from BUMPT cells transfected with plasmid of MBD2 plus with or without EGR1 neutralizing antibody for 24 h, which indicated by the co-culture model diagram of BUMPT cells and murine embryonic NIH 3T3 fibroblasts. (H) Immunoblot analysis of FN, Col I and IV, and α-SMA. (I) Analysis of the gray-scale image between them. (J) Concentration of EGR1 by ELISA. Data are expressed as means ± SD (n = 6). #p < 0.05 versus scramble or control group. ∗p < 0.05 versus TGF-β1 or MBD2 group.