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. 2016 Jul 21;11(7):e0159682. doi: 10.1371/journal.pone.0159682

Fig 4. Tauroursodeoxycholic acid (TUDCA) prevents cardiac fibrosis in HeartCRT+ mice.

Fig 4

(A) Trichrome staining for collagen deposition in the myocardium of control, transgenic and HeartCRT+ mice fed TUDCA. The arrows indicate the location of blue staining for collagen. (B) Quantitative analysis of the percentage of area with collagen deposition in control (ctrl), and HeartCRT+ animals fed tamoxifen (TAM) or TAM+ TUDCA at day 14 and day 21 *p<0.01. Data are representative of 6 biological replicates. (C) Immnunoblot analysis and quantification of TGFβ1 protein in hearts from HeartCRT+ mice fed TAM or TAM+TUDCA. Antibodies to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were used as a loading control. Data are representative of 6 biological replicates. (D) Real-time Q-PCR analysis for TGFβ1 transcript in hearts from HeartCRT+ animals fed TAM or TAM+ TUDCA. * p = 0.0254 ** p = 0.0148. Data are representative of 6 biological replicates. TAM samples were normalized to Day 0 and TAM+TUDCA samples were normalized to the corresponding day in TAM treatment to highlight the TUDCA effect. (E) Abundance of collagen type 1A1 in wild-type (wt) and transgenic mice fed TAM+TUDCA for 14 or 21 days. Antibodies to human influenza hemagglutinin (HA) were used to probe for the abundance of HA-tagged recombinant calreticulin (CRT). Anti-GAPDH antibodies were used as a loading control. (F) Abundance of collagen 1A1 mRNA in hearts from HeartCRT+ animals fed TAM or TAM+TUDCA. NS, not significant. *p = 0.0424. Data are representative of 6 biological replicates. TAM samples were normalized to Day 0 and TAM+TUDCA samples were normalized to corresponding day in TAM treatment to highlight the TUDCA effect. (G) Western blot analysis of periostin in hearts from TAM+TUDCA fed mice. Anti-HA antibodies were used to probe for abundance of HA-tagged recombinant calreticulin (CRT). Anti-GAPDH antibodies were used as a loading control. (H) Abundance of periostin mRNA in hearts from HeartCRT+ animals fed TAM or TAM+TUDCA. Data are representative of 6 biological replicates. TAM samples were normalized to Day 0 and TAM+TUDCA samples were normalized to corresponding day in TAM treatment to highlight the TUDCA effect. 3–5 animals were used for each analysis.