Fig. 4

Nalbuphine inhibits EMT and metastasis. a–c MDA-MB-231 and MCF-7 cells were treated with nalbuphine for 48 h and levels of indicated mRNA and protein were determined by RT-PCR (a, b) and western blot (c) (n = 3). d MDA-MB-231 cells were treated with nalbuphine for 48 h and then subjected to wound healing analysis, representative images (left) and statistical analysis (right) are shown (n = 3). e MDA-MB-231 cells were treated with nalbuphine for 48 h and migration and invasion were then determined by transwell assays. Representative images (left) and statistical analysis (middle and right) are shown (n = 3). f Primary cells were extracted from Ctrl and Nal tumors. Levels of indicated proteins were determined by western blot (n = 5). g Primary cells were extracted from Ctrl and Nal tumors. Levels of indicated proteins were determined by western blot (n = 5). Data are presented as mean ± SEM. p-value was determined by Student’s t-test (*p < 0.05, **p < 0.01, ***p < 0.001). See also Additional file 5: Figure S4