FIGURE 5 from MARCH2, a Novel Oncogene-regulated SNAIL E3 Ligase, Suppresses Triple-negative Breast Cancer Metastases

PTK6 inhibition enhances MARCH2:SNAIL association. A, MDA-MB-231 cells expressing FLAG-Snail and HA-MARCH2 (WT) were treated with vehicle control or P21d for 4 or 18 hours in the presence of MG132. MARCH2–SNAIL interaction was assessed by immunoblotting FLAG immunoprecipitates with anti-HA antibody. B, Endogenous MARCH2–SNAIL interaction following P21d treatment of MDA-MB-231 was assessed by Duolink PLA. Cells were treated with DMSO or P21d for 8 hours and MG132 cotreatment for the last 4 hours. Cells were stained with antibodies to Snail and MARCH2. PLA signals/cell were quantitated using ImageJ (left). Scale bar, 15 µm.