Fig 2: GV1001 suppresses TGF-ß signaling and EMT in NHOKs exposed to IR. NHOKs were exposed to 6 Gy IR and maintained in culture for 10 days in the presence of GV1001 (1 µM) or TRI (1 µM). Western blot analysis was performed for TGF-ß signaling molecules, p-Smad2/3 and Smad4, and TGF-ß target mesenchymal markers, ZEB1, FN, N-Cad and Snail. GAPDH was used as a loading control. NHOKs, normal human oral keratinocytes; IR, ionizing radiation; TGF-ß, transforming growth factor-ß; TRI, TGF-ß receptor inhibitor; Smad, small mothers against decapentaplegic; p-, phosphorylated; ZEB1, zinc finger E-box binding homeobox 1; FN, fibronectin; N-Cad, N-Cadherin.

Fig 3: Ufbp1 deficiency resulted in nuclear translocation of Smad3. (A) Subcellular localization of p-Smad3. (B) Nuclear and cytoplasmic extracts were blotted for pSmad3 and Smad4 of MEFs. (C) Ufbp1 overexpression affected Smad3 phosphorylation. (D) Interaction of Ufbp1 with Smad2 and Smad3 in co-immunoprecipitation assays, using IgG served as a negative control. Data are presented as means ± SEM, ***P < 0.001 (n = 3). Scale bar = 20 µm.

Fig 4: SLC39A5 depletion suppresses the TGF-ß signalling. (A) show the intracellular zinc level of the knockout (KO) cells was reverted after SLC39A5 re-expression via lentivirus infection. (B,C) show the rescue of expression of the extracellular matrix members (COL1, FN and LN) and Smad proteins (Smad1, pSmad1/5/9, Smad2/3, pSmad2/3 and Smad4) in the knockout (KO) cells after SLC39A5 re-expression via lentivirus infection. (D–G) show the degradation of cycloheximide (CHX)-treated Smad1, Smad2/3 and Smad4 at different time points within 8 h. Degradation curves (E–G) were obtained based on statistical analyses of the Western blotting results (D). *p < 0.05, **p < 0.01 and ***p < 0.001

Fig 5: Effect of the combination of G-Rh2 and L-OHP on LoVo/L-OHP cells. LoVo cells were treated with or without 15 µmol/ml L-OHP, or 15 µmol/ml L-OHP + 250 µg/ml G-Rh2 for 24 h. (A) A 3-(4,5 dimethylthiazol-z-yl)-3,5-diphenyltetrazolium bromide assay was used to detect cell viability, and (B) flow cytometry was used to detect cell apoptosis. (C) Protein levels of P-gp, Smad4, Bcl-2, Bax, and caspase-3 in LoVo cells were determined using a western blot assay. mRNA levels of (D) P-gp, (E) Smad4, (F) Bcl-2, (G) Bax and (H) caspase-3 were determined by reverse transcription-quantitative polymerase chain reaction analysis. Data are expressed as the mean ± SD. **P<0.01 vs. control group. G-Rh2, ginsenoside Rh2; L-OHP, oxaliplatin; P-gp, P-glycoprotein.