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. Author manuscript; available in PMC: 2016 Sep 28.
Published in final edited form as: Dev Cell. 2015 Sep 10;34(6):642–655. doi: 10.1016/j.devcel.2015.08.014

Figure 6. MAP4K1/2/3/5 is required for Hpo/Mst-independent phosphorylation and nuclear exclusion of YAP induced by F-actin disruption.

Figure 6

(A) Concomitant depletion of all four MAP4Ks leads to elevation in the expression levels of three Hpo pathway target genes ctgf, cyr61, and survivin in the Mst1/2 null HCCs. Total RNAs were prepared from HCCs transfected with the indicated siRNAs and the relative mRNA levels of indicated genes were measured by qPCR (mean±SD, n=3). Note the significant increase of gene expression when all four kinases were depleted. ** denotes a p-value<0.01.

(B) Concomitant depletion of all four MAP4Ks promotes the proliferation of Mst1/2 null HCCs.

(C) Depletion of MAP4K1/2/3/5 by RNAi reduces LatB-induced YAP and Lats phosphorylation in the Mst1/2 null HCCs. HCCs cultured on fibronectin-coated surface at low cell density were treated with 1μg/mL LatB for 1 hr and then analyzed by western blotting. Note that LatB induced YAP and Lats phosphorylation (compare lanes 1 and 2). Both inductions were diminished when four kinases were depleted (compare lanes 2 and 6).

(D–E) Depletion of MAP4K1/2/3/5 by RNAi attenuates LatB-induced nuclear exclusion of YAP in the Mst1/2 null HCCs. HCCs transfected with control siRNAs or siRNAs against MAP4K1/2/3/5 were cultured on chamber slides coated with fibronectin at low cell density. Cell were treated with 1μg/mL LatB for 1 hr and then fixed and stained with YAP antibody, phalloidin for F-actin, and DAPI for cell nuclei (C). 100 cells were quantified for YAP localization and the results were shown in (D).

See also Figure S5.