Fig. 2: The paracrine effect of rCAFs, but not sCAFs, promotes cancer cell chemoresistance.

a, b DDP IC₅₀ experiments of FaDu or Tu686 cells after co-culture with or without sCAFs or rCAFs (n = 3 independent experiments). c, d Co-culturing GFP overexpressing FaDu or Tu686 cells with RFP expressing sCAFs or rCAFs for 48 h. Bar charts represent the relative number of cancer cells or CAFs in each group (n = 3 independent samples). Each result is shown after being normalized to the control in each experiment. e, f Migration assays of FaDu or Tu686 cells co-culture with sCAFs or rCAFs. Bar charts present the relative number of migrated cells in each group. g, h Invasion assays of FaDu or Tu686 cells co-cultured with sCAFs or rCAFs (n = 3 independent samples). i, j DDP IC₅₀ experiments of FaDu or Tu686 cells in the presence or absence of conditioned medium (CM) from sCAFs or rCAFs. Untreated cancer cells were used as a control (ctrl). k, l Colony formation assays of FaDu or Tu686 cells after treated with/without CM from sCAFs or rCAFs. m, n Bar charts show the relative migrated cell number in each group. o, p Invasion assays of FaDu and Tu686 cells after exposed with CM from sCAFs or rCAFs. q, r Representative bright field images of tumor spheres from each group are given. Bar charts show the relative sphere area in each group (n = 3 independent samples). s Schematics diagram depicting the DDP treatment strategy of mice bearing tumors derived from subcutaneous injection of FaDu cells together with/without sCAFs or rCAFs at 1 to 3 ratio. t Representative gross tumor image from each treatment group is given. u Line graph shows the tumor growth of each group (n = 5 mice per group). v Bar chart shows the mean of tumor weight in each group. Data are shown as means ± S.E.M. *P < 0.05, **P < 0.01, ***P < 0.001, N.S. non-significant. c–r, v One-way ANOVA. u Two-way ANOVA. Scale bars in (c–h, m–r) represent 100 μm, (t) 1 cm.