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. 2021 Jul 1;22(13):7145. doi: 10.3390/ijms22137145

Figure 2.

Figure 2

The effects of ANO1 inhibition on HaCaT cells. (a) Proliferation of cells exposed to 30 µM T16Ainh-A01 for 3 days. (b) Proliferation of cells exposed to 10 µM Ani9 for 3 days. (c) Proliferation of cells following ANO1 knock-down for 3 days. After knock-down of ANO1 using siRNA targeting ANO1 and in negative controls (mock and si-NC), proliferation rates of HaCaT cells were measured for 3 days. (d) Immunoblots of ANO1 and ERK and AKT signaling pathway-related proteins after ANO1 knock-down for 2 days. (e) Densitometric analysis for the relative intensity of ANO1 and ERK and AKT signaling pathway-related proteins. Treatments with the two blockers (T16Ainh-A01 and Ani9) and ANO1 knock-down were performed as four independent replicates to guarantee reliable results. To compare the differences between control (vehicle or Mock [si-NC]) and blocker-treated or ANO1 knock-down cells, two-way ANOVA and Tukey’s post-hoc tests were used. *: significantly different from vehicle in (a,b), from Mock and si-NC (c), and from Mock (e) (* p < 0.05, ** p < 0.01, and *** p < 0.001). #: significantly different from si-NC (e) (# p < 0.05 and ## p < 0.01).