TFEB and TFE3 are novel components of the integrated stress response

A

Venn diagram showing overlapping genes in analyzed data sets.

B, C

Schematic representations of TFE3 binding region in the ATF4 promoter in MEF cells treated with tunicamycin (0.1 μg/ml) for 16 h (B) or starved in EBSS for 2 h (C). The distance upstream of the transcription start site (TSS) of a potential E‐box sequence is indicated in base pairs (bp).

D, E

E‐box distance from transcription start site for targets categorized as lysosomal genes in MEF treated with tunicamycin (0.1 μg/ml) for 16 h (D) or starved in EBSS for 2 h (E). Peaks called as TFE3 binding sites were analyzed for E‐box elements using UCSC genome browser and distance from transcription start site was assessed. Graphs represent the number of genes with E‐box distance × (in kbp) from the transcription start site.

F

Relative quantitative real‐time PCR analysis of SUMF1, DERL1, ATF6, APEX1, and GPX1 mRNA transcript levels in ARPE‐19 cells infected with either adenovirus expressing Null or TFEB‐S211A (mean ± SD of the RNA fold change of indicated mRNAs normalized to actin mRNA from three independent experiments, Student's t‐test, **P < 0.01, ***P < 0.001).

G

Immunoblots of protein lysates from ARPE‐19 cells infected with either adenovirus expressing Null or TFEB‐S211A. Data are representative of three independent experiments.

H

Quantification of IRE1α and APEX1 protein levels in ARPE‐19 cells infected as indicated in (G) (mean ± SD of the fold increase of the indicated protein to actin ratio from three independent experiments, Student's t‐test, ***P < 0.001).

Figure 7. TFEB and TFE3 regulate expression of critical regulators of the ER stress response.