Figure 5. Deletion of MAP4K4/6/7 in MST1/2 dKO cells abolishes the majority of LATS and YAP phosphorylation induced by various signals.
(a) Western blots showing CRISPR-mediated deletion of MST1/2 and MAP4K4/6/7 (MM-5KO) in HEK293A cells. (b) Density-induced LATS and YAP phosphorylation is significantly compromised in MM-5KO HEK293A cells. S, sparse, 1.5 × 105 cells per well were seeded onto six-well plates 24 h before collecting. D, Dense, 8.0 × 105 cells per well were seeded. (c) Energy stress-induced LATS and YAP phosphorylation is abolished in MM-5KO HEK293A cells. (d) Serum deprivation-induced LATS and YAP phosphorylation is largely blocked in MM-5KO HEK293A cells. (e) Actin depolymerization-induced YAP phosphorylation is largely blocked in MM-5KO HEK293A cells. (f) YAP/TAZ are constitutively localized in nucleus in MM-5KO cells even under serum deprivation for 1 h. Scale bar, 10 μm. (g) Quantification of percentage of the cells with more nuclear (N) or cytosolic YAP/TAZ (C) signals was performed in three randomly chosen fields for each treatment. Typically, each field contains 80–150 cells.