Fig 2: Specific cell-surface marker genes of invasive fibroblasts promoted invasion and fibrosis. (A) Cell sorting strategy of F3, SEMA7A, and ITGA6 negative and high fibroblasts for following experiments. (B and C) Relative mRNA levels (B; n = 4 per group) and total protein levels (C) of F3, SEMA7A, and ITGA6 expression in sorted F3, SEMA7A, and ITGA6 negative and high fibroblasts. (D and E) Representative images (D) and index (E; SEMA7ANeg/High migration/invasion, n = 4 per group; F3Neg/High migration, n = 12 per group, invasion, n = 9 per group; ITGA6Neg/High migration, n = 9 per group, invasion, n = 12 per group) of migration and invasion of SEMA7A, F3, and ITGA6 negative and high fibroblasts. (F) Cell adhesion of SEMA7A and ITGA6 high and negative fibroblasts was quantified (n = 4 per group). (G) Overexpression of SEMA7A was confirmed by Western blotting. (H) Cell-surface expression of SEMA7A in SEMA7A overexpression and control fibroblasts. (I and J) Representative images (I) and index (J; n = 3 per group) of migration and invasion of SEMA7A overexpression fibroblasts. (K) Quantification of percentage of SEMA7A+ fibroblasts by flow cytometry on freshly isolated normal and IPF human lungs (Normal, n = 4; IPF, n = 7). (L and M) Trichrome staining (L) and hydroxyproline (M; n = 10 per group) of mice lungs receiving SEMA7A high and negative fibroblasts, and age-matched mice were treated with culture medium only. Dash-boxed regions were shown at higher magnification. CTL, control; OE, overexpression. Scale bar: 1 mm (D and I), 500 µm (L). Three or four independent experiments were performed on fibroblasts from different patients (B, E, F, J, K, and M). Data are the mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 by Student’s t test (B, E, F, and I–K) and two-way ANOVA (M). Source data are available for this figure: SourceData F2.

Fig 3: KDM5B inhibitor GSK‐467 blocks HCC progression in vitro and in vivo. A, Cell sphere formation evaluated by cell sphere formation assay. B, Colony formation ability evaluated by clonogenic assay. C, Cell migration detected by scratch test. D, Cell invasion detected by Transwell assay. E, Weight of transplanted tumours. F, Growth curve of subcutaneous transplanted tumours. G, RT‐qPCR to detect the expression of KDM5B, miR‐448, YTHDF3 and ITGA6 in tumour tissues. H, Western blot analysis to detect the expression of KDM5B, YTHDF3 and ITGA6 in tumor tissues. I, Representative images showing xenografts in nude mice. J, Immunohistochemistry of KDM5B, YTHDF3 and ITGA6 proteins in tumour tissues (500×). The data in multiple groups were compared using one‐way ANOVA and Tukey's post hoc test. The data in groups at different time were analysed by repeated measures ANOVA followed by Bonferroni's post hoc test. *P < .05 when compared with the NS group. & P < .05 when compared with the GSK‐467 + oe‐NC group

Fig 4: KDM5B activates the YTHDF3/ITGA6 axis through suppressing miR‐448 to promote HCC progression. Cells transfected with oe‐NC + sh‐NC, oe‐KDM5B + sh‐NC or oe‐KDM5B + sh‐ITGA6 were subcutaneously planted into nude mice. A, Weight of xenograft tumours after different treatments. B, Growth curve of subcutaneous xenograft tumours after different treatments. C, RT‐qPCR determination of KDM5B, miR‐448, YTHDF3 and ITGA6 expression in tumour tissues after different treatments. D, Western blot analysis of KDM5B, YTHDF3 and ITGA6 expression in tumour tissues normalized to GAPDH after different treatments. E, Representative images showing xenograft tumours in nude mice. F, Immunohistochemistry of KDM5B, YTHDF3 and ITGA6 proteins in tumour tissues (500×). The data were measurement data, which were presented as mean ± standard deviation. The data in multiple groups were compared using one‐way ANOVA and Tukey's post hoc test. The data in groups at different time were analysed by repeated measures ANOVA followed by Bonferroni's post hoc test. *P < .05 when compared with treatment of oe‐NC + sh‐NC. & P < .05 when compared with treatment of oe‐KDM5B + sh‐NC

Fig 5: miR-448 targets YTHDF3/ITGA6 axis. A, Venn diagram predicting downstream genes of miR-448 based on TargetScan, StarBase, mirDIP, miRDB and microRNA. B, Box plot of YTHDF3 expression after obtained by StarBase, wherein the left red boxes indicate its expression in HCC samples, and the right purple boxes represent its expression in normal samples. C, Box plot of ITGA6 expression obtained by StarBase, wherein the left red boxes represent its expression in HCC samples, and the right purple boxes represent its expression in normal samples. D, Expression correlation diagram between YTHDF3 and ITGA6 in HCC obtained by StarBase. E, MEM analysis displaying a significant co-expression relationship between YTHDF3 and ITGA6. F, Binding sites between miR-448 and YTHDF3. G, Luciferase activity after cotransfection of miR-448 mimic and pmirGLO-YTHDF3-WT or pmirGLO-YTHDF3-MUT relative to Renilla luciferase activity. H, RT-qPCR determination of mRNA level of YTHDF3 and ITGA6 in HCC cells after miR-448 mimic and miR-448 inhibitor treatment. I, Western blot analysis of the protein level of YTHDF3 and ITGA6 in HCC cells normalized to GAPDH after miR-448 mimic and miR-448 inhibitor treatment. J, Detection of YTHDF3 and ITGA6 mRNA expression in HCC cells after cotransfection of sh-KDM5B and miR-448 inhibitor by RT-qPCR. K, Detection of YTHDF3 and ITGA6 protein expression in HCC cells after cotransfection of sh-KDM5B and miR-448 inhibitor by Western blot analysis. The data were measurement data, which were presented as mean ± standard deviation. The data in two groups were compared using unpaired t test, and the data in multiple groups were compared using one-way ANOVA and Tukey's post hoc test. *P < .05 when compared with treatment of sh-NC