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. 2017 Jul 3;36(17):2510–2528. doi: 10.15252/embj.201696089

Figure 7. The YAP‐Skp2 axis does not seem to be conserved in mice.

Figure 7

  1. Immunoblots of mouse or human cell lines transduced with vector‐ or 5SA‐YAP‐encoding retroviruses. Although 5SA‐YAP expression induces the expression of Ctgf, the best‐known Yap target gene, in both mouse and human cells, it only induces Skp2 expression in human cells.
  2. Immunoblot of Lats1−/− Lats2fl/fl MEFs infected with the indicated shRNA lentiviruses and transduced with or without Cre. The specificity of the antibody for mouse Skp2 was validated using Skp2 knockdown.
  3. Immunoblot of mouse cell lines transfected with the indicated siRNAs for 2 days.
  4. (Above) Experimental protocol schematic. (Below) immunoblots of MDA‐MB‐231 (left) or 4T1 cells (right) treated with 100 ng/ml nocodazole (Noc) 30 h after siRNA transfection (siC: siControl, siY: siYAP) to arrest them in mitosis. Mitotic cells were then collected via a mechanical shake‐off. As a control, non‐transfected mitotic mock (Mo) cells were washed with PBS several times and released into early G1 (Noc. Rel.).
  5. Immunoblot of 4T1 cells transfected with the indicated siRNAs for 2 days and then treated with MG132 for 4 h.
  6. Immunoblots of vector‐ or 5SA‐YAP‐expressing AML12 mouse cells placed in suspension for the indicated times.
  7. A sequence alignment of the region of the Skp2 promoter surrounding the TB2 TEAD‐binding consensus site for several mammalian species.
  8. TEAD‐binding motifs −5,000 to +5,000 bp from the TSS in the Skp2 promoter of the indicated species.
  9. Immunoblots of MDCK cells (a dog cell line) transduced with either vector‐ or 5SA‐YAP‐encoding retroviruses.