Figure 5
From: A miR-130a-YAP positive feedback loop promotes organ size and tumorigenesis
The YAP-miR-130a-VGLL4 positive feedback loop mediates Hippo signaling. (A, B) Knockdown of YAP, TAZ and TEADs increases VGLL4 protein level. Cells were transfected with siRNAs (A) or infected with shRNA (B) and analyzed by immunoblotting. Cyr61 is a known YAP target and a positive control. (C) miR-130a blocks VGLL4 induction upon YAP and TAZ inhibition in HMLE cells. (D) The miR-130a sponge abrogates VGLL4 inhibition by YAP in HepG2 cells. (E) Knockdown of Lats1/2 represses VGLL4 protein level. Cells were transfected with siRNAs as indicated and analyzed by immunoblotting. (F) Inactivation of the Hippo pathway in vivo increases miR-130a and represses VGLL4 protein level. Control and albumin-Cre-mediated Mst1/2 conditional knockout livers were harvested at 2 months of age for analysis. (G) Loss of cell adhesion represses miR-130a in a Lats1/2-dependent manner. siRNA-transfected HMLE cells were cultured in adhesion “A” or in suspension “S” for 48 h for analysis of miR-130a level. (H, I) Cell suspension induces VGLL4 by inhibiting YAP and miR-130a. HMLE stable cells were transfected and cultured in adhesion or in suspension for 48 h and analyzed by immunoblotting. (J) miR-130a inhibits anoikis in cooperation with YAP. HMLE stable cells were subjected to suspension culture for 48 h. Anoikis rates were determined by Annexin V staining and FACS. (K) Inhibition of miR-130a promotes anoikis. HepG2 cells were transfected and analyzed as in J.
