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. 2022 Mar 1;18(10):2333–2349. doi: 10.1080/15548627.2022.2029671

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This work was authored as part of the Contributor’s official duties as an Employee of the United States Government and is therefore a work of the United States Government. In accordance with 17 U.S.C. 105, no copyright protection is available for such works under U.S. Law.

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Figure 5. — The FACT complex promotes TFEB-dependent expression of oxidative stress genes. (A) Relative quantitative real-time PCR analysis of PPARGC1A, FOXO3, HIF1A, MAFF, MAFG, HMOX1, PYROXD1, TXNRD1, SRXN1, ADNP2 and SESN2 mRNA transcript levels in HeLa WT cells infected with adenovirus expressing TFEB-S211A-Flag compared to Null for 40 h. Data represented as geometric mean ± SD and significance tested with Student’s t-test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001) from at least three independent experiments. (B) Immunoblot analysis of protein lysates from HeLa WT cells infected with either Null adenovirus or adenovirus expressing TFEB-S211A-Flag for 40 h. (C) Quantification of immunoblots shown in (B). Data represents mean relative protein levels ± SD with n = 3. Significance tested using Student’s test (*p < 0.05, **p < 0.01, ***p < 0.001). (D) Relative quantitative real-time PCR analysis of PPARGC1A, HIF1A, MAFF, MAFG, HMOX1 and ADNP2 mRNA transcript levels in HeLa WT cells infected with adenovirus expressing TFEB-S211A-Flag upon incubation with curaxin for 4 h at different doses. Significance tested with two-way ANOVA with Sidak’s multiple comparisons test (**p < 0.01, ***p < 0.001, ****p < 0.0001) from three independent experiments. (E) Schematic representations of the TFEB-Flag binding region in the promoter of PPARGC1A, HIF1A, MAFF, HMOX1 and ADNP2 in HeLa-TFEB-Flag cells transfected with either control or FACT siRNAs treated with NaAsO₂ (50 μM) for 6 h. The transcription start site is indicated as TSS.