Fig. 3: Mechanical compression regulates multiple cellular responses in a Hic-5-dependent manner.

A Representative Western blots demonstrate the effective knockdown (KD) of Hic-5 in bronchial epithelial cells at baseline and in response to mechanical compression. Compression induced Hic-5 expression in Hic-5 wildtype (WT) cells, whereas this Hic-5 induction was abolished in Hic-5 KD cells. Densitometry analysis was performed, and the fold change relative to GAPDH was calculated in arbitrary units (A.U.) (mean ± SD, n = 5 HBE cell donors). P values: one-way ANOVA with Holm–Šídák post hoc correction. Uncropped versions of the blots are provided as a Source Data file. B, C Volcano plots present differentially expressed genes (DEGs) between control and compressed conditions (DESeq2 test, two-sided). In Hic-5 WT cells, mechanical compression induced a significant number of DEGs (B). In Hic-5 KD cells, the number of DEGs were significantly fewer compared to WT cells (C). n = 3 HBE cell donors. D The volcano plot presents DEGs between Hic-5 WT and Hic-5 KD cells under compression. In B–D, upregulated genes are marked by red, downregulated genes by green, and not significant genes by gray. E The top 10 DEGs were ranked by Log₂ (fold change) between Hic-5 WT vs Hic-5-KD under compression. F Gene Ontology (GO) analysis identified biological processes and molecular functions of upregulated genes in Hic-5 WT cells compared with Hic-5 KD under compression, as demonstrated in (D). G The enrichment network visualized functional modules of upregulated genes in Hic-5 WT cells compared with Hic-5 KD under compression, as demonstrated in (D).