Fig 2: KDM3A promotes transcription of a pro-fibrotic gene program. a Volcano plot of differentially expressed genes in Kdm3a KO mouse hearts compared to those in WT mouse hearts at 6 weeks post-TAC surgeries. Timp1 is among the most downregulated genes in KO hearts. b Heatmaps of differentially expressed genes in KO vs WT and Kdm3a-Tg vs WT mouse hearts at 6 weeks post-TAC surgery, showing differentially expressed genes are inversely regulated in KO vs Tg hearts. c GO analysis of differentially downregulated genes in KO vs WT TAC mouse hearts. Relative mRNA of genes involved in fibrosis in Kdm3a-/- (d) and Kdm3a-Tg (e) mouse hearts 6 weeks post-Sham and TAC surgery. mRNA transcripts were measured by qRT-PCR, normalized against internal Gapdh, and expressed relative to Sham WT mice. n = 5 ± SEM. *, #p < 0.05 (ANOVA). *, WT TAC vs. WT Sham. #, Tg-TAC vs. WT-TAC

Fig 3: Confirmation of hub genes. (A) qPCR test of 15 hub genes compared with RNA-seq result. The trend of fold change were similar between RNA-seq and qPCR; (B) IF staining of protein PF4, which is encoded by hub gene Pf4; (C) IHC staining of protein C3, CCL6, CCR2, PAFr, and TIMP1, which are coded by hub genes C3, Ccl6, Ccr2, Ptafr, and Timp1,(×400).

Fig 4: JIB-04 blocks TAC-induced hypertrophic remodeling and fibrosis. a Experimental protocol of JIB-04 and control vehicle (Veh) treatment of WT and Kdm3a-Tg mice. JIB-04/Veh were given to mice at either day 1 or 2 (exp 1) or 21 (exp 2) post-TAC. b %FS, c HW/BW ratio, d relative cell size, e percentage of fibrotic area, and f relative fold change of fetal genes and Timp1 of WT sham, WT TAC and Kdm3a-Tg TAC mouse heart at the end of experiment 1. n = 5-6 ± SEM, *, #, $, p < 0.05 (ANOVA). *, WT TAC vs WT Sham. #, JIB-04 WT TAC vs Veh WT-TAC. $, JIB-04 Tg TAC vs Veh TG TAC. g Western blot of TIMP1 in vehicle or JIB-04 treated WT mouse hearts at post-sham/TAC week 6 in experiment 1. h %FS over treatment period, i HW/BW, j relative cell size, and k % fibrotic area at end of experiment 2 of Sham and TAC Kdm3a-Tg mouse hearts treated with vehicle or JIB-04. n = 5 ± SEM, *p < 0.05 (ANOVA)

Fig 5: The mRNA (A) and protein (B) expressions of TIMP-1, TIMP-2, MMP-2 and MMP-9in human oral squamous carcinoma HSC-3 cells. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. the control group. Geniposide-L: 25 µg/mL geniposide; LcS-geniposide-L: 1.0 × 106 CFU/mL LcS + 25 µg/mL geniposide; Geniposide-H: 50 µg/mL geniposide; LcS-geniposide-H: 1.0 × 106 CFU/mL LcS + 50 µg/mL geniposide.