Fig. 5

Inhibition effect of Rg3 on CAFs formation and activation. A Concentration of TGF-β in 4T1-cultured medium (CM) after the treatment of PBS, DTX, C-Lp/DTX, Nanoxel-PM, Rg3, Rg3-Lp, Rg3/DTX and Rg3-Lp/DTX, respectively. (n = 3) B Western blot detection of α-SMA and GADPH on 3T3 cells after treated with TGF-β (20 ng/ml), TGF-β/SB-431542, PBS, different conditioned 4T1 medium and 4T1-CM/SB-431542. C Western blot detection of α-SMA, p-Smad2/3 and GADPH on 3T3 cells treated with different conditioned 4T1 medium. D IF observation over α-SMA in 3T3 cells after treatment with TGF-β (20 ng/ml), TGF-β/SB-431542, PBS, 4T1-CM, 4T1-CM /SB-431542 and different conditioned 4T1 medium. E ELISA assay of the concentration of TGF-β in tumor tissues after the treatment of PBS, DTX, C-Lp/DTX, Nanoxel-PM, Rg3, Rg3-Lp, Rg3/DTX and Rg3-Lp/DTX, respectively. (n = 3) F q-PCR assay of α-SMA level in tumor tissues after the treatment of PBS, DTX, C-Lp/DTX, Nanoxel-PM, Rg3, Rg3-Lp, Rg3/DTX and Rg3-Lp/DTX, respectively. (n = 3) G Western blot detection of p-Smad2/3, α-SMA, β-actin and GADPH in tumors after treatment with PBS, DTX, C-Lp/DTX, Nanoxel-PM, Rg3, Rg3-Lp, Rg3/DTX and Rg3-Lp/DTX, respectively. H Flow cytometry analysis of activated CAFs in tumor after treatment with PBS, DTX, C-Lp/DTX, Nanoxel-PM, Rg3, Rg3-Lp, Rg3/DTX and Rg3-Lp/DTX, respectively. (n = 3) *p < 0.05, **p < 0.01, and ***p < 0.001; Differences between two groups were analyzed with unpaired t-test, one-way ANOVA was performed to compare data among multiple groups; Error bars represent mean ± SD of three independent experiments