Fig. 5: ENSA activates STAT3 to promote the transcription of SREBP2 in a PP2A-dependent manner. | Nature Communications

Fig. 5: ENSA activates STAT3 to promote the transcription of SREBP2 in a PP2A-dependent manner.

From: Copy number amplification of ENSA promotes the progression of triple-negative breast cancer via cholesterol biosynthesis

Fig. 5

a JASPAR prediction of STAT3-binding sites on the sequence of SREBP2. b qRT-PCR and PCR analysis of STAT3 at the SREBP2 promoter after ChIP assays in MDA-MB-231 cells expressing control or ENSA shRNA. n = 3. Data are presented as mean ± SD. Two-tailed unpaired Student’s t tests. c qRT-PCR detecting relative SREBP2 mRNA expression in BT549 and MDA-MB-231 cells after STAT3 transient silencing. n = 3. Data are presented as mean ± SD. Two-tailed unpaired Student’s t tests. d Luciferase reporter assay detecting the activity of the SREBP2 promoter in BT549 cells ± ENSA knockdown and ± STAT3 overexpression. n = 3. Data are presented as mean ± SD. Two-tailed unpaired Student’s t tests. e Western blotting images showing the expression of enzymes involved in cholesterol biosynthesis, SREBP2, pSTAT3-Tyr705 and total STAT3 in BT549 and MDA-MB-231 cells with ± ENSA knockdown and ± STAT3 overexpression. n = 3 independent experiments. f Filipin III staining showing the cellular free cholesterol contents of BT549 and MDA-MB-231 cells ± ENSA knockdown and ± STAT3 overexpression. n = 3 independent experiments. g Western blotting images showing the expression of STAT3, pSTAT3-Tyr705, and pSTAT3-Ser727 in MDA-MB-231 cells expressing control or PPP2CA siRNA. n = 3 independent experiments. h Western blotting images showing the expression of pSTAT3-Tyr705 and SREBP2 in MDA-MB-231 cells ± ENSA knockdown and ± transient PPP2CA knockdown. n = 3 independent experiments. Source data are provided as a Source Data file.

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