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. 2023 Oct 24;97:104846. doi: 10.1016/j.ebiom.2023.104846

Fig. 4.

Fig. 4

Functional validation of identified pro-tumoral fibroblasts. (a) Kaplan–Meier curves regarding the overall survival of myoCAFs (upper) and vCAFs (lower) based on their gene signature in all CC cohort from the TCGA database. P-value was calculated by Log-rank test. (b–c) mIF staining of myoCAFs (upper: POSTN+/COL1A1+) and vCAFs (lower: LMCD1+/COL1A1+) in collected CSCC (n = 32) and CAde (n = 11) samples. ∗P < 0.05, ∗∗∗P < 0.001. The P-values were calculated by Wilcoxon Rank Sum Test. (d) Immunofluorescence staining of myoCAFs (upper: POSTN+/COL1A1+) and vCAFs (lower: SOD2+/COL1A1+) sorted by flow cytometry from one clinical CC sample. (e–f) Transwell migration assay of CC cells treated with different condition medium from primary myoCAFs and vCAFs, respectively. Transwell assay had three independent biological replicates. ∗∗∗P < 0.001. The P-values were calculated by Wilcoxon Rank Sum Test. (g) Blood vessel formation assay of HUVEC treated with different condition medium from primary myoCAFs, vCAFs or Con group. ∗P < 0.05, ∗∗P < 0.01, Ns not significant. Blood vessel formation assay had three independent biological replicates. The P-values were calculated by Wilcoxon Rank Sum Test. (h) mRNA expression levels of some marker genes in primary myoCAFs and vCAFs. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001. Each target gene had three independent biological replicates. The P-values were calculated by Wilcoxon Rank Sum Test.