Fig. 4: Nonsecretory ECM1b inhibits myosin phosphorylation and tumor growth. | Nature Communications

Fig. 4: Nonsecretory ECM1b inhibits myosin phosphorylation and tumor growth.

From: Extracellular matrix protein-1 secretory isoform promotes ovarian cancer through increasing alternative mRNA splicing and stemness

Fig. 4

a Colocalization of integrin αX or β2 with HA (ECM1a/1b) in HeyA8 cells treated with medium A and B collected from A8i-A and A8i-B cells, respectively (bars = 10 µm). b Coomassie Brilliant Blue (R-250) staining of co-IP products prepared with cell lysates (CLs) of A8i-A and A8i-B cells and an anti-HA antibody after separation by SDS-PAGE; the arrows indicate the protein bands analyzed by mass spectrum (MS). c Results of secondary ion MS analyses of the most abundant proteins in A1 and B1 samples derived from ECM1a and ECM1b, respectively. d Direct binding of ECM1a or ECM1b to nonphosphorylated and phosphorylated myosin IIa detected by co-IP and WB analysis using corresponding antibodies. e Colocalization of ECM1b with nonphosphorylated myosin IIa detected by IF in cells (bar = 10 µm). Expression of myosin IIa and phosphorylated myosin IIa detected by WB analysis in ECM1-silenced cells and ECM1a- and ECM1b-overexpressing cells (f), in WT and MT ECM1-overexpressing cells (g, the actin loading control in g was also used for the same samples in Fig. 3f left panel because these tests were parallelly performed), in integrin αX- or integrin β2-knockdown cells (h), and in ECM1a cells (A8i-A) transfected with ECM1b (A8i-A + B) or ECM1b cells (A8i-B) transfected with ECM1a (A8i-B + A) (i).

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