Skip to main content
. 2023 May 12;24(10):8676. doi: 10.3390/ijms24108676

Figure 4.

Figure 4

TGF-β neutralizing antibody does not affect caALK5-mediated effects on the metastatic outgrowth in liver of B16F10 cells. (A) Analysis of the protein expression of TGF-β signaling components by Western blot of B16F10 caALK5 cells treated with dox, TGF-β3, SB505124 or TGF-β-neutralizing antibody 1D11. Vinculin level was used as a loading control. (B) Analysis of gene expression by qPCR in B16F10 caALK5 cells treated without or with dox (1 μg/mL), TGF-β3 (1 ng/mL), SB505124 (1 μM) or TGF-β-neutralizing antibody 1D11 treatment (10 μg/mL). Effects of caALK5, TGF-β and 1D11 treatment is measured by mRNA levels of Serpine1, Ctgf, and Smad7. Gene expression was corrected by the housekeeping genes Gapdh and Hprt. Mean ± S.D., technical duplicates. Representative of 3 biological repeats. (C) Schematic representation of the experimental set-up of in vivo experiment. Mice were injected with B16F10 or B16F10 caALK5 cells and were treated with control chow or dox-containing chow (625 mg/kg) for two weeks. Mice were injected with IgG control or 1D11 antibody (5 mg/kg), 6 mice per condition. (D) Examples of livers containing B16F10 or caALK5 expressing B16F10 liver metastasis, treated with IgG or 1D11. (E) Metastatic liver replacement was calculated for 4 sections per liver of mice from in vivo experiment shown in (C,D). n = 24, mean ± S.E.M. ** p ≤ 0.01, two-way ANOVA.