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. 2022 Oct 7;13:5919. doi: 10.1038/s41467-022-33521-4

Fig. 7. S100A4 increases ZIP1 expression in fibroblasts.

Fig. 7

a, b ZIP1 expression in mCAF treated with DOX at the indicated doses (a) or for the indicated times (b). Representative results from three (a, b) independent experiments are shown. c S100A4 levels in the same volume of LLC-GFP-luc cell culture mediums (CMs) with or without DOX treatment determined by western blotting. The same number of tumour cells were seeded and pre-treated with DOX (1 μM) for 6 h. Twenty-four hours after changing the fresh medium, CMs were collected. The same volume of CM from different treatment groups was analysed by Western blotting. A representative result from three independent experiments is shown. d ZIP1 expression in mCAF treated with indicated CM. For S100A4 neutralisation, CM was pre-incubated with α-S100A4 antibody 3B11 (6 μg/mL) for 1 h. A representative result from four independent experiments is shown. e, f ZIP1 expression in mCAF treated with S100A4 at the indicated doses (e) or for the indicated times (f). β-actin was used as control. The samples derived from the same experiment and blots were processed in parallel. Representative results from three (e, f) independent experiments are shown. g, h Expression of phosphorylated p65 (pp65), p65, phosphorylated ERK (pERK) (g), phosphorylated AKT (pAKT), AKT, phosphorylated p38 (pp38), and p38 (h) in mCAF treated with S100A4 for the indicated time periods. β-actin was used as control. The samples derived from the same experiment and blots were processed in parallel. Representative results from three (g, h) independent experiments are shown. i Expression of ZIP1 in mCAF treated with S100A4, SB202190 (p38 inhibitor, 50 μM), U0126 (ERK inhibitor, 10 μM), SC75741 (p65 inhibitor, 8 μM), and LY294002 (AKT inhibitor, 25 μM) as indicated. A representative result from three independent experiments is shown. j Expression of ZIP1 in mCAF treated with S100A4, TLR4-IN-C34 (TLR4 inhibitor, 10 μM), and FPS-ZM1 (RAGE inhibitor, 1 μM), as indicated. A representative result from three independent experiments is shown. k Diagram showing the signalling pathways for the regulation of ZIP1 by S100A4. l Extracellular S100A4 in LLC-GFP-luc tumours, with or without DOX treatment. n = 6 for the PBS group and n = 7 for the DOX group. Data are presented as mean ± SEM, n = 6 for PBS, n = 7 for DOX. Two-tailed t-test. Source data are provided as a Source Data file (a–j, l).