Fig. 2: Inhibiting miR-3202 protects against DUX4-induced cell death by upregulating FAIM2. | Cell Death & Disease

Fig. 2: Inhibiting miR-3202 protects against DUX4-induced cell death by upregulating FAIM2.

From: Antiapoptotic Protein FAIM2 is targeted by miR-3202, and DUX4 via TRIM21, leading to cell death and defective myogenesis

Fig. 2

A Cell viability indicated by CellTitre-Glo luminescence, measuring total ATP content in control non-transfected, non-induced, and scramble- or miR-3202 inhibitor-transfected doxycycline-induced LHCN-M2iDUX4 cells. ****P < 0.0001 by one-way ANOVA with Tukey’s post hoc test (n = 5). The experiment has been repeated five independent times. B Cell viability of iDUX4 rhabdosarcoma cells, transfected and induced as in A. ****P < 0.0001, ***P = 0.0002 by one-way ANOVA with Tukey’s post hoc test (n = 5). C Cell viability of 293TiDUX4 cells, transfected and induced as in A. Data are presented as individual data points plus mean; ****P < 0.0001, **P = 0.0065 by one-way ANOVA with Tukey’s post hoc test (n = 5) the experiment has been repeated 3 independent times. D FACS analysis of FAM-labeled miR-3202 inhibitor transfection efficiency across the duration of the assay, showing over 90% transfection efficiency in LHCN-M2iDUX4 cells. E Western blot for DUX4 expression in LHCN-M2iDUX4 cells transfected (or not) with miR-3202 inhibitor for 24 h and induced (or not) with 50 ng/ml doxycycline for another 24 h. F Western blot for DUX4 and FAIM2 expression in LHCN-M2iDUX4 cells transfected with miR-3202 inhibitor or controls, with or without subsequent induction of DUX4 by 50 ng/ml doxycycline. Far-right lane shows LHCN-M2iDUX4 cells transfected with FAIM2-expressing plasmid. G Cell viability Assay of LHCN-M2iDUX4 cells induced with 50 ng/ml for 48 h following transfection with FAIM2 expression plasmid versus a control non-coding plasmid. ****P < 0.0001, by one-way ANOVA with Tukey’s post hoc test (n = 5).

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