Supplementary Figure 3: Muc5ac promotes epigenetic alterations in CAF precursor cells

<p dir="ltr">Supplementary Figure 3: Muc5ac promotes epigenetic alterations in CAF precursor cells. (A) Expression profile of DNA methyltransferases (Dnmt) and ten-eleven translocation dioxygenases (Tet) in AD-MSCs treated with Muc5ac-proficient or Muc5ac-deficient conditioned media (CM). (B) Expression profile of DNMT and TET family genes in fibroblasts from high- and low-Muc5ac–expressing (based on ductal expression) patients from single-cell RNA-seq dataset CRA001160. (C) Expression levels of histone H3 lysine 27 acetylation (H3K27Ac) and total histone H3 in PSCs, AD-MSCs, and murine cancer-associated fibroblasts (CAFs) derived from KPC tumors. (D) H3K27Ac levels in murine PSCs treated with increasing concentrations of CM derived from KPC-3266 cells (Muc5ac-expressing). (E) H3K27Ac expression in PSCs treated with KPC-3266 CM, heat-inactivated CM (Hi), CM with CXCR2 inhibitor (iC2), CM with RAC1 inhibitor (iR1), or left untreated (C). (F) H3K27Ac levels in PSCs after treatment with KPC-3266 (P) and KPC-3266 M5KO CM (M5KO) and heat-inactivated CM (Hi). (G) Differential expression of significantly modulated histone methyltransferases (HMTs), histone demethylases (HDMs), and histone deacetylases (HDACs) from RNA-seq analysis of PSCs treated with Muc5ac-proficient or Muc5ac-deficient CM for 5 days. (H) qRT-PCR analysis of Hdac4 expression in PSCs treated with Muc5ac-proficient or Muc5ac-deficient CM. For all gene expression data, expression was normalized to β-actin. For western blot densitometry involving H3K27Ac, quantification was normalized to total histone H3. Data represents mean ± standard deviation (SD) from three independent biological replicates. Statistical analysis was performed using an ordinary one-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.0001.</p>