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. 2022 Nov 7;24(1):e54042. doi: 10.15252/embr.202154042

Figure EV2. Relationships among HIF‐1, p53, and ZBTB2.

Figure EV2

  • A, B
    The indicated cells transfected with p5HRE‐Luc were transiently co‐transfected with pEF6/ZBTB2 (ZBTB2), pcDNA4/UCHL1 (UCHL1), pcDNA4/IDH3α (IDH3α), pcDNA4/Ly6E (Ly6E), or each of their corresponding control vectors (EV), pEF6/myc‐His B, pcDNA4/myc‐His A, pcDNA4/myc‐His A, or pcDNA4/myc‐His A, respectively. The cells were then cultured under the indicated oxygen conditions for 24 h and subjected to the luciferase assay.
  • C
    The indicated cells were treated with scramble‐siRNA (Scr) or ZBTB2‐siRNA (siZBTB2), cultured under < 0.1% oxygen conditions for 24 h, and subjected to qRT–PCR for ZBTB2 mRNA levels.
  • D
    The indicated cells transiently transfected with p53‐Luc (#219083, Agilent Technologies) were additionally transfected with pcDNA3/p53 R175H, R248W, R273H, or pcDNA3 (EV) as indicated, cultured under < 0.1% oxygen conditions for 24 h, and subjected to the luciferase assay.
  • E
    The indicated cells transiently transfected with either pGL3/(3′Gli‐BS)4‐Luc or its empty vector, pGL3‐Luc (EV), were additionally transfected with either pEF6/ZBTB2 (ZBTB2) or pEF6/myc‐His B (EV), cultured 20% oxygen conditions for 24 h, and subjected to the luciferase assay.

Data information: pRL‐CMV was used as an internal control in every luciferase assay (A, B, D, E). Mean ± s.d. The number of technical replicates in all of the experimental groups was 3, and the reproducibility of the results was confirmed at least three times by biologically independent experiments (A–E). N.S., not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, Student's t‐test.