Fig. 1: SIX1 promotes transcription, translation, and secretion of IL6 to promote proliferation, migration, and invasion of breast cancer cells.
From: SIX1 transmits signals for breast cancer progression via the ZEB1/IL6/STAT3 signaling axis
A RT-qPCR analysis of IL6 mRNA levels in 66cl4 transfected with SCR or shSix1 and MCF7 transfected with OE Ctrl or OE SIX1, n = 3. B RT-qPCR analysis of IL6 mRNA levels in indicated groups, n = 3. C RT-qPCR analysis of SIX1 mRNA levels in indicated groups, n = 3. D Western blot analysis of SIX1 and IL6 protein levels in Six1-knockdown 66cl4, followed by IL6 overexpression. E Quantitation of Western blot in (D), n = 4. F Western blot analysis of SIX1 and IL6 protein levels in SIX1 overexpressed MCF7, followed by IL6 knockdown. G Quantitation of Western blot in (F), n = 4. H ELISA analysis of IL6 content in the cellular supernatant of Six1-knockdown 66cl4, followed by IL6 overexpression, n = 3. I ELISA analysis of IL6 content in the cellular supernatant of SIX1 overexpressed MCF7, followed by IL6 knockdown, n = 3. J Proliferation curves of Six1-knockdown 66cl4, followed by IL6 overexpression, n = 4. K Proliferation curves of SIX1 overexpressed MCF7, followed by IL6 knockdown, n = 4. L Representative migration assay (0–24 h) on Six1-knockdown 66cl4, followed by IL6 overexpression, scale bar is 100 μm. M Quantitation of cell migration in (L), n = 3. N Representative invasion assay on Six1-knockdown 66cl4, followed by IL6 overexpression, scale bar is 50 μm. O Quantitation of cell invasion in (N), n = 3. P Representative IF of E-cadherin (red) and N-cadherin (green) in SIX1 overexpressed MCF7, followed by IL6 knockdown. DAPI (blue), scale bar is 20 μm. Q Representative IF of Vimentin (pink) in SIX1 overexpressed MCF7, followed by IL6 knockdown. DAPI (blue), scale bar is 20 μm. R Quantitation of relative fluorescence intensity in (P) and (Q), n = 4. Data were presented as means ± SD; ANOVA (B, C, E, G, H, I, M, O, R, and the left panel of A) or two-sided Student’s t-test (the right panel of A) were used for statistical analysis. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, and ns P > 0.05. SCR scramble, OE Ctrl overexpressing control, shNT shRNA of negative control, sh#1 #2 #3 three independent shRNA constructs targeting genes.
