Fig 6. Knockdown of TRPM4 prevents cell death caused by H₂O₂ challenge in H9c2 cardiomyocytes.

(A) Quantitative RT-PCR confirming the gene silencing of TRPM4. siNEG, cells transfected with control siRNA; siTRPM4, cells transfected with TRPM4-targeting siRNA. n = 5 for each group. (B) Confirmation of suppressed TRPM4 protein expression by immunocytochemistry 48 h after siRNA transfection. Green, anti-TRPM4, Blue, Hoechst 33342 dye (nuclei). The fluorescent images were overlaid with DIC images of the cultures. (C) Confirmation of suppressed TRPM4 protein expression by Western blot 24 h after siRNA transfection. (D) Impact of gene silencing on the loss of viability induced by 200 μM H₂O₂. Cell viability was measured by the MTT assay. n = 5 for each group. (E) Impact of TRPM4 knockdown on the hypoxia/reoxygenation (H/R) challenge. Cell viability was measured by the MTT assay. n = 6 for each group. Statistical analysis was performed using Dunnett’s test as post hoc. *: p < 0.05, **: p < 0.01, N.S.: p > 0.05.