Figure 2: Six1 is necessary (downstream of Snail1/Twist1) and sufficient to mediate NCA effects. | Nature Communications

Figure 2: Six1 is necessary (downstream of Snail1/Twist1) and sufficient to mediate NCA effects.

From: EMT cells increase breast cancer metastasis via paracrine GLI activation in neighbouring tumour cells

Figure 2

(a) Western blot analyses performed on WCLs from HMLER-Ctrl, HMLER-Snail1 and HMLER-Twist1 cells transfected with 150 nM of siSix1 or a non-targeting small interfering RNA (siRNA) pool (siNT). HDAC1, loading control. (b,c) Representative 7–8 h migration assay of HMLER-Ctrl cells in CM from HMLER-Snail1 or Twist1 cells ±siSix1. (d,e) Quantification of cell migration in b,c. (f) Western blot analyses performed on WCLs from MCF7-Ctrl and MCF7-Six1 cells cultured in indicated CM for 48 h. β-Tub, β-Tubulin, loading control; CK18, cytokeratin 18; FN1, fibronectin. (g) Representative ICC of E-Cad (red) in MCF7-Ctrl and Six1 cells cultured in indicated CM for 48 h (DAPI, blue). Quantification of % membranous E-Cad, n≥100; scale bar, 20 μm. (h) Representative anoikis resistance graph (plotted as absorbance as a measure of cell number) of MCF7-Ctrl or MCF7-Six1 cells cultured in indicated CM for 24 h. C-cells, M-Media; Scale bar, 100 μm, s.e.m. shown, n≥3, one-way ANOVA with Tukey’s post test in all cases; NS, not significant; *P<0.05, **P<0.01 and ***P<0.001.

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