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. 2022 Apr 8;7(7):e152330. doi: 10.1172/jci.insight.152330

Figure 5. Piezo1 mediates mechanical stretch– or compression-induced profibrotic responses in HK2 cells.

Figure 5

(A) mRNA level of Piezo1 in HK2 cells pretreated with 5 μM GsMTx4 followed by cyclic stretch. Data are shown as mean ± SEM (n = 6 in each group). (B and C) Representative immunoblots and corresponding densitometry analysis of Piezo1 protein abundance in HK2 cells pretreated with 1 μM or 5 μM GsMTx4 followed by cyclic stretch. Data are shown as mean ± SEM (n = 4 in each group). (D) Fluo-4-AM was added to HK2 cells transfected with Piezo1 siRNA followed by cyclic stretch, and the fluorescence signals of calcium were detected by flow cytometry analysis. (E and F) Representative immunoblots and corresponding densitometry analysis of fibronectin, α-SMA, and E-cadherin protein abundance in HK2 cells pretreated with 5 μM GsMTx4 followed by cyclic stretch. Data are shown as mean ± SEM (n = 4 in each group). *P < 0.05 when compared with static and #P < 0.05 when compared with stretch by 1-way ANOVA with Student-Newman-Keuls test. (G and H) Representative immunoblots and corresponding densitometry analysis of fibronectin, α-SMA, and E-cadherin protein abundance in HK2 cells transfected with Piezo1 siRNA followed by cyclic stretch. Data are shown as mean ± SEM (n = 4 in each group). *P < 0.05 when compared with scramble siRNA static and #P < 0.05 when compared with scramble siRNA stretch by 1-way ANOVA with Student-Newman-Keuls test. (I and J) Representative immunoblots and corresponding densitometry analysis of Piezo1, fibronectin, and α-SMA protein abundance in HK2 cells pretreated with 5 μM GsMTx4 followed by 15 mmHg compression for 24 hours. Data are shown as mean ± SEM (n = 4 in each group). *P < 0.05 when compared with CTL and #P < 0.05 when compared with compression by 1-way ANOVA with Student-Newman-Keuls test.