Fig. 4: MetR suppresses fibrotic gene expression by targeting Hoxc8.

A Motif analysis of transcription factor (TF) binding sites in TGF-β-upregulated genes in NRK-49F cells. B Heatmap showing TFs sensitive to MetR treatment in NRK-49F cells. C Quantitative RT-PCR analysis of mRNA levels for Hoxc8, Hoxc9, Prrx2, and Hoxb9 in the indicated groups. From left to right: ****P = < 0.0001, ns P = 0.1239, **P = 0.0047, ***P = 0.0006, respectively, by two-tailed unpaired Student’s t-test. Data are presented as mean ± SEM. D Quantitative RT-PCR analysis of mRNA levels for fibrosis marker genes (Acta2, Col1a1, and Col3a1) in NRK-49F cells transfected with control siRNA or siRNAs targeting Hoxc8, Prrx2, and Hoxb9. ****P = < 0.0001. by two-tailed unpaired Student’s t-test. Data are presented as mean ± SEM. E Western blot analysis of Hoxc8 in NRK-49F cells treated with TGF-β and MetR. F Western blot analysis of Hoxc8, α-SMA, Col1a1, and Fibronectin expression in NRK-49F cells treated with TGF-β and siHoxc8. G Heatmap showing gene expression values in NRK-49F cells treated with or without TGF-β and siHoxc8. Rows represent Z-scores calculated for each group. H Venn diagram illustrating the overlap between MetR-suppressed and siHoxc8-suppressed genes (left panel). GO analysis of the overlapping genes (right panel). I Quantitative RT-PCR analysis of mRNA levels for fibrosis marker genes, Vim and Acta2, in NRK-49F cells with or without Hoxc8 overexpression under the indicated conditions. From left to right: *P = < 0.0417, ns P = 0.6504, **P = 0.0016, ns P = 0.2471, respectively, by two-tailed unpaired Student’s t-test. Data are presented as mean ± SEM. J Western blot analysis of α-SMA and Hoxc8 expression in NRK-49F cells with or without Hoxc8 overexpression and MetR treatment. Independent experiments = 3. Source data are provided as a Source Data file.