Empagliflozin inhibits angiotensin II-induced hypertrophy in H9c2 cardiomyoblasts through inhibition of NHE1 expression

Fig. 1. ANG-induced increase in cell surface area of H9c2 cardiomyoblasts was reduced in the presence of EMPA. Left: Representative images of H9c2 cardiomyoblasts stained with crystal violet. Right: Cell surface area represented as mean value ± SEM of n = 4 independent experiments (n = 3 for EMPA and ANG + EMPA groups). # indicates P < 0.01 vs CTRL. β indicates P < 0.01 vs ANG

Fig. 2. A ANG-induced SGLT-1 protein expression was reduced in the presence of EMPA in H9c2 cardiomyoblasts. Upper: Representative Western Blot of H9c2 cardiomyoblasts cell lysate probed with anti-SGLT-1. Tubulin was used as loading control. Lower: Quantification of SGLT-1 band normalized to tubulin. The bar graph represents the mean value ± SEM of n = 5 independent experiments (n = 3 for EMPA, and ANG + EMPA).* indicates P < 0.05 vs CTRL. # indicates P < 0.01 vs CTRL. α indicates P < 0.05 vs ANG. B SGLT-2 protein expression of H9c2 cardiomyoblasts with ANG ± EMPA treatment. Representative Western blot of H9c2 cardiomyoblasts cell lysate probed with anti-SGLT-2. Tubulin was used as loading control

Fig. 3. ANG-induced expression of NHE1 was reduced by EMPA in H9c2 cardiomyoblasts. Upper: Representative Western blot of H9c2 cardiomyoblasts cell lysate probed with anti-NHE1. Tubulin was used as loading control. Lower: Quantification of NHE1 band normalized to tubulin. The bar graph represents the mean value ± SEM of n = 4 independent experiments (n = 3 for ANG, and ANG + EMPA). * indicates P < 0.05 vs control. α indicates P < 0.05 vs ANG

Fig. 4. ANG, EMPA, or their combination does not affect the phosphorylation of p90 ribosomal s6 kinase protein expression H9c2 cardiomyoblasts. Upper: Representative Western blot of H9c2 cardiomyoblasts cell lysate probed with anti-phospho p90 RSK and RSK2. Tubulin was used as loading control. Lower: Quantification of p90 RSK normalized to RSK2 and tubulin. The bar graph represents the mean value ± SEM of n = 3 independent experiments

Fig. 5. ANG, EMPA, or their combination does not affect the phosphorylation of Akt protein expression H9c2 cardiomyoblasts. Upper: Representative Western blot of H9c2 cardiomyoblasts cell lysate probed with anti-phospho-Akt. Tubulin was used as loading control. Lower: Quantification of phospho-Akt normalized to tubulin. The bar graph represents the mean value ± SEM of n = 4 independent experiments (n = 3 for EMPA, and ANG + EMPA)