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. 2020 Mar 9;21(4):e48389. doi: 10.15252/embr.201948389

Figure 1. Identification of HOXA4 as a negative regulator of YAP/TEAD transcriptional activity through a pooled shRNA screen.

Figure 1

  • A
    Dynamic fluorescence patterns according to cell density in 8×GTIIC‐EmGFP 293T cells.
  • B
    EmGFP intensity in LATS2 knockdown or forced expression of a constitutively active form of YAP (YAP‐S127A). Scale bars indicate 100 μm.
  • C
    Schematic drawing of FACS‐based pooled shRNA screen of molecules that affect the YAP/TEAD signaling pathway in 8×GTIIC‐EmGFP 293T cells.
  • D
    Scatter plots of mean frequency of each shRNA per gene. HOXA4 is indicated as a red dot.
  • E
    Augmented 8×GTIIC‐luciferase reporter activity by loss of HOXA4. ShSAV1 was used as an experimental control. **P < 0.01 versus shCtrl, by ANOVA with Sidak's correction.
  • F, G
    Quantitative real‐time PCR analysis of ANKRD1 expression in HEK 293T cells transfected with shHOXA4 (F) or HOXA4 expression vector (G). **P < 0.01, by unpaired two‐tailed Student's t‐test.
Data information: Graphs of (E–G) show mean ± SEM; three biological repeats.