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. 2018 Apr 12;37(11):e97780. doi: 10.15252/embj.201797780

Figure EV6. Inhibition of YAP/TAZ induces senescence‐like phenotypes.

Figure EV6

  1. WI38 cells transfected with the indicated siRNAs: control (siCo.) or YAP/TAZ (siYT) and examined for gene expression of established YAP/TAZ target genes. n = 4.
  2. Representative EdU stainings and nuclear counterstains (DAPI) of WI38 cells quantified in Fig 7B. Scale bar: 80 μm.
  3. Western blot analysis for p‐ERK levels in WI38 cells depleted of YAP/TAZ. GAPDH is a loading control.
  4. qPCR for SASP genes on MDA‐MB‐231 cells transfected with control (siCo.) or YAP/TAZ (siYT) and collected at 2 days. n = 4.
  5. Quantification of MDA‐MB‐231 cells transfected as in (C) and stained for SAβgal activity 4 days after transfection. Representative results of a single experiment (> 200 cells per condition); n = 4.
  6. qPCR on WI38 cells plated as in Fig 7F–I on soft hydrogels, and examined for expression of established YAP/TAZ target genes. n = 4.
  7. Quantification of proliferation (EdU stainings) on WI38 cells plated on fibronectin‐coated glass coverslips (stiff) or fibronectin‐coated hydrogels (soft). Representative result of a single experiment (> 100 cells per condition); n = 4.
  8. Quantification of proliferation (EdU stainings) on WI38 cells plated on fibronectin‐coated glass coverslips (Unpatt.), on medium‐small (2,025 μm2) or on small (1,024 μm2) fibronectin‐coated small islands. Representative result of a single experiment (> 100 cells per condition); n = 2.
  9. Representative immunofluorescence images of YAP/TAZ in WI38 cells plated on fibronectin‐coated glass coverslips (Unpatt.) or on small (1,024 μm2) microprinted fibronectin‐coated islands. DAPI serves as nuclear counterstain. Scale bar: 15 μm.
  10. WI38 cells were infected with control vector (−) or YAP 5SA‐encoding retrovirus, treated with 100 ng/ml or 20 ng/ml doxorubicin and assayed by EdU staining. Representative result of a single experiment (> 100 cells per condition); n = 4.
  11. WI38 cells were infected with control vector (−) or YAP 5SA‐encoding retrovirus, treated with 100 ng/ml doxorubicin and stained for SAβgal activity. Representative result of a single experiment (> 100 cells per condition); n = 4.
Data information: Data are mean and SD with *P < 0.05 (two‐tailed Student's t‐test).