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. Author manuscript; available in PMC: 2022 Sep 1.
Published in final edited form as: Mol Cancer Res. 2022 Mar 1;20(3):373–386. doi: 10.1158/1541-7786.MCR-21-0208

Figure 1. Upregulation of multiple GPCRs promotes breast cancer EMT.

Figure 1.

A-C, Effect of stable AT1R expression in ZR75–1 cells on cell morphology (A), expression of EMT markers by immunoblot analysis (B), and plasma membrane expression of both E- and N-cadherin species by immunofluorescence staining and confocal microscopy (C). Scale bar, 5 μm. D and E, Effect of stable PAR1 expression in MCF7 cells on cell morphology (D) and expression of EMT markers (E). F, Impact of siRNA-mediated AT1R or PAR1 knockdown in BT549 and MDA-MB-231 cells, respectively, on EMT markers. G, Effect of stable AT1R expression in ZR75–1 cells on cell migration, as measured continuously over time. A representative time-course is shown at left, with blue pseudocolor mask highlighting the progressive migration of cells into scratch wounds placed at time 0 hrs. Quantification of scratch would closure is shown at right, plotted as a continuous function of time (mean ± SD, n=10), ****, P < 0.0001, two-way ANOVA. H, Effect of stable AT1R expression in ZR75–1 cells on invasiveness, as measured using matrigel-coated Boyden chambers. Representative images of invaded cells are shown at left. Quantification of cell invasion is shown at right (mean ± SEM, n=13), ****, P < 0.0001, two-tailed t test with Welch’s correction. I, Effect of stable PAR1 expression in MCF7 cells on cell migration, using the same scratch wound assay described in panel G (mean ± SD, n=12), ****, P < 0.0001, two-way ANOVA. J, Effect of stable PAR1 expression in MCF7 cells on invasiveness, using the same matrigel invasion assay described in panel H (mean ± SEM, n=15), ***, P < 0.001, two-tailed t test with Welch’s correction.