Fig. 7

CRABP2 mediates ubiquitination of Lats1 in breast cancer cells dependent on ER status. a Expression of CRABP2, ER, Lats1, E-cadherin, ZO-1 and Vimentin protein was examined in T47D and MCF7 cells expressing CRABP2 (CRABP2) and vector (Con), with or without ER (si-ER) by Western blotting. b Expression of CRABP2, ER, Lats1, E-cadherin, ZO-1 and Vimentin protein was examined in BT549 and MDA-MB-231 cells expressing CRABP2 siRNA (si-CRABP2) and siRNA (Con), with or without ER (ER) by Western blotting. c The ubiquitination of Lats1 was examined in ER⁺ breast cancer cells (T47D cells) stably expressing CRABP2 shRNA (sh-CRABP2) and shRNA (sh-NC). Ubiquitin-HA and Flag-Lats1 were transfected into sh-NC and sh-CRABP2 T47D cells. d The ubiquitination of Lats1 was examined in ER⁻ breast cancer cells (BT549 cells) expressing CRABP2. Ubiquitin-HA, CRABP2 and Flag-Lats1 were transfected into BT549 cells. e The ubiquitination of Lats1 was examined when ER was knocked down and CRABP2 was overexpressed in T47D cells at the same time. Various combinations of si-ER, CRABP2 and Flag-Lats1 and ubiquitin-HA were transfected into T47D cells. f The ubiquitination of Lats1 was examined when ER was overexpressed and CRABP2 was knocked down in BT549 cells at the same time. Various combinations of ER, si-CRABP2 and Flag-Lats1 and ubiquitin-HA were transfected into BT549 cells. Ubiquitination Lats1 was tested by IP of Lats1 with anti-Flag antibody, and analyzed by anti-HA antibody with Western blotting