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. 2022 Feb 28;12(3):281. doi: 10.3390/membranes12030281

Figure 5.

Figure 5

Monitoring of TRPV4-mediated cell death in HCEC-12 (12) and its derivatives, 12EV and 12V4. Morphology of HCEC-12- and HA-TRPV4-transduced HCEC-12 (a) either untreated (left) or treated with the TRPV4 antagonist RN1734 (middle) or the TRPV4 agonist GSK101 (right). Differential interference contrast (DIC) micrographs; scale bar 100 µm. Metabolic activity of GSK101- (b) or RN1734- (c) treated cells were analyzed by resazurin conversion assay. Cells were treated with GSK101 (d) or RN1734 (e), and membrane integrity was quantified by flow cytometry after vital staining with SYTOX Green to assess cell death. Triangles (▲) represent statistically significant differences compared to the untreated control sample, HCEC-12 (12); squares (◼) represent statistically significant differences compared to untreated HCEC-12EV (12EV); circles (●) represent statistically significant differences compared to untreated HCEC-12V4 (12V4) and asterisks (*) represent statistically significant differences between the untreated cell populations. Data are presented as the means ± SEM of n = 36 from four independent experiments (b,c) or n = 16 from two independent experiments (d,e). * p < 0.01, ** p < 0.001 and *** p = < 0.0001, one-way ANOVA with Tukey’s multiple comparisons test.