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. 2022 Mar 14;16(9):1891–1912. doi: 10.1002/1878-0261.13200

Fig. 4.

Fig. 4

NOX4 regulates TGFβ1‐induced stem cell‐related proteins. (A) U3031MG cells were transiently transfected with control (siControl) or NOX4 (siNOX4, pool of four siRNA sequences) siRNAs and stimulated with TGFβ1 for 24 h, mRNA expression levels analysed by qPCR, data represent the mean ± SEM (n = 5 independent experiments and each with technical triplicate); statistics: two‐way ANOVA test, Tukey’s multiple comparison. (B) mRNA expression levels analysed by qPCR: U3031MG or U3034MG cells were stimulated with TGFβ1 for 24 h in with or without NOX1/4 inhibitor. Data represent the mean ± SEM (n = 2 independent experiments and each with technical triplicate); statistics: two‐way ANOVA test, Tukey’s multiple comparison. (C) U3031MG cells were transiently transfected with control (siControl) or NOX4 (siNOX4, pool of four siRNA sequences) siRNAs and stimulated with TGFβ1 for 24 h, immunoblot of the indicated proteins, Gapdh or β‐actin, are used as a loading control. (C) A representative experiment of (D) Quantification of immunoblot data using the densitometric analysis of each protein, normalised with total protein using Stain‐Free precast gels using image lab™ software (Bio‐Rad, Sundbyberg, Sweden); data represent mean ± SEM (n = 3–6 independent experiments, depending on the antibody), statistics: two‐way ANOVA test, Sidak’s multiple comparison. (A, B, D) Statistical comparison indicates *P < 0.05, **P < 0.01, ***P < 0.001.