Journal: Journal of Cellular and Molecular Medicine

Article Title: PU.1 inhibition attenuates atrial fibrosis and atrial fibrillation vulnerability induced by angiotensin‐II by reducing TGF‐β1/Smads pathway activation

doi: 10.1111/jcmm.16678

Figure Lengend Snippet: Inhibition of PU.1 in mice down‐regulated the Ang‐II‐induced activation of the atrial tissue and atrial fibroblasts TGF‐β1/Smads pathway in vivo and in vitro. (A) Representative Western blots and (B–D) quantitative densitometric analyses showing the expression of TGF‐β1, p‐Smad3 and p‐Smad2/3 in the atrial tissue induced by the subcutaneous infusion of Ang‐II with or without DB1976 (n = 5). (E) Representative Western blots and (F–H) quantitative densitometric analyses showing the expression of TGF‐β1, p‐Smad3 and p‐Smad2/3 in the cultured atrial fibroblasts sourced from Ang‐II‐induced mice and treated with and/or Ang‐II/DB1976 for 24 h (n = 3). Data represent the mean ± SD. * P < .01 vs. the sham/control group, # P < .05 vs. the Ang‐II group. GAPDH was used as the internal control. Ang‐II, angiotensin‐II

Article Snippet: Mice in the sham and Ang‐II groups were given saline, while mice in the sham+DB1976 group and Ang‐II+DB1976 group were administered with DB1976 (5 mg/day/kg; Glpbio, Montclair, CA, USA) by intraperitoneal injection once a day for 28 days, as previously described.

Techniques: Inhibition, Activation Assay, In Vivo, In Vitro, Western Blot, Expressing, Cell Culture, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: PU.1 inhibition attenuates atrial fibrosis and atrial fibrillation vulnerability induced by angiotensin‐II by reducing TGF‐β1/Smads pathway activation

doi: 10.1111/jcmm.16678

Figure Lengend Snippet: Inhibition of PU.1 in mice attenuated the atrial fibroblast proliferation and differentiation induced by Ang‐II in vivo and in vitro. (A) Representative immunofluorescence images of PCNA expression in the atrial tissue of mice induced by the subcutaneous infusion of Ang‐II. Red staining indicates PCNA expression. Nuclei were counterstained with DAPI. Scale bar, 20 µm. (B, D) Representative Western blots and (C, E) quantitative densitometric analyses showing the expression of PCNA and α‐SMA in the atrial tissue inducted by Ang‐II in vivo (n = 5). (F‐N) Atrial fibroblasts sourced from Ang‐II‐induced mice treated with and/or Ang‐II/DB1976 for 24 h. (F, J) Representative Western blots and (G, K, L, M, N) quantitative densitometric analyses showing the expression of PCNA, α‐SMA, ED‐A Fn, Smemb and DDR2; (H) Absorbance values of the CCK‐8 proliferation assay; (I) representative immunofluorescence images of α‐SMA expression (red staining). Scale bar, 20 µm. (O‐W) Atrial fibroblasts sourced from Ang‐II‐induced mice treated with and/or Ang‐II/si‐PU.1 for 24 h. (O, Q, S) Representative Western blots and (P, R, T, U, V, W) quantitative densitometric analyses showing the expression of PU.1, PCNA, PCNA, α‐SMA, ED‐A Fn, Smemb and DDR2 (n = 3). Data represent the mean ± SD. * P < .01 vs. the sham/control group. # P < .05 vs. the Ang‐II group/Ang‐II + si‐Con group. GAPDH was used as the internal control. Ang‐II, angiotensin‐II; CCK‐8, Cell Counting Kit‐8; DAPI, 4′,6‐diamidino‐2‐phenylindole

Article Snippet: Mice in the sham and Ang‐II groups were given saline, while mice in the sham+DB1976 group and Ang‐II+DB1976 group were administered with DB1976 (5 mg/day/kg; Glpbio, Montclair, CA, USA) by intraperitoneal injection once a day for 28 days, as previously described.

Techniques: Inhibition, In Vivo, In Vitro, Immunofluorescence, Expressing, Staining, Western Blot, CCK-8 Assay, Proliferation Assay, Control, Cell Counting

Journal: Journal of Cellular and Molecular Medicine

Article Title: PU.1 inhibition attenuates atrial fibrosis and atrial fibrillation vulnerability induced by angiotensin‐II by reducing TGF‐β1/Smads pathway activation

doi: 10.1111/jcmm.16678

Figure Lengend Snippet: Inhibition of PU.1 in mice attenuated the atrial fibrosis induced by Ang‐II in vivo and in vitro. (A) Representative images of Masson's trichrome staining of the interstitial, sub‐epicardial and perivascular regions of the atrial tissue. Blue staining indicates the fibrotic tissue. Scale bar, 20 µm. (B) Quantitative analysis of fibrosis (n = 5). (C) Representative Western blot and (D, E) quantitative densitometric analyses showing the expression of CTGF and collagen I in the atrial tissue induced by the subcutaneous infusion of Ang‐II (n = 5). (F, I) Representative Western blots and (G, H, J, K) quantitative densitometric analyses showing the expression of CTGF and collagen I in the cultured atrial fibroblasts sourced from the Ang‐II‐induced mice and treated with or without Ang‐II, DB1976 or si‐PU.1 for 24 h (n = 3). Data represent the mean ± SD. * P < .01 vs. the sham/control group, # P < .05 vs. the Ang‐II group/Ang‐II + si‐Con group. GAPDH was used as the internal control. Ang‐II, Angiotensin‐II; si‐PU.1, small interfering RNA against PU.1

Article Snippet: Mice in the sham and Ang‐II groups were given saline, while mice in the sham+DB1976 group and Ang‐II+DB1976 group were administered with DB1976 (5 mg/day/kg; Glpbio, Montclair, CA, USA) by intraperitoneal injection once a day for 28 days, as previously described.

Techniques: Inhibition, In Vivo, In Vitro, Staining, Western Blot, Expressing, Cell Culture, Control, Small Interfering RNA