Fig. 5

Neuregulin (NRG1) induces jagged 1 (JAG1) expression in macrophages and is important for in vitro transendothelial migration (iTEM). a Western blot (left) showing induction of JAG1 protein expression in BAC after NRG1 stimulation. Quantitation of protein level is shown on the right. Data are means and SEM from three independent experiments; *p < 0.001, t test. b Quantitation of JAG1 mRNA induction in BACs after co-culture with MDA-MB 231. Values expressed as fold change of co-culture versus BAC alone. Data are means and SEM trom three independent experiments; **p < 0.001 by z test. c Western blot of clustered regularly interspaced short palindromic repeats (CRISPR) knockout of JAG1 in BAC cells showing lack of JAG1 expression compared to control guide RNA (gRNA) in the presence or absence of NRG1. d Transendothelial migration of MDA-MB 231 cells alone, with control gRNA BACs, or JAG1 gRNA knockout BACs, confirming the importance of macrophage JAG1 in intravasation. Data are means and SEM from three independent experiments; *p < 0.05, t test. e Transendothelial migration of BT549 cells alone, with control gRNA BACs, or JAG1 gRNA knockout BACs, further confirming the importance of macrophage JAG1 in intravasation. Data are means and SEM from three independent experiments; **p < 0.01, t test. f Transendothelial migration of MDA-MB 231 cells in the presence of a scrambled control peptide (scramble) or JAG1 peptide (Jag1). Data are means and SEM from three independent experiments; *p < 0.05, paired t test