Fig. 2: Loss of Kindlin-2 inhibits AR transcriptional activity and its downstream events. | Cell Death & Disease

Fig. 2: Loss of Kindlin-2 inhibits AR transcriptional activity and its downstream events.

From: Kindlin-2 promotes Src-mediated tyrosine phosphorylation of androgen receptor and contributes to breast cancer progression

Fig. 2Fig. 2

A Representative immunofluorescence staining of AR (green) in control (Ctrl siRNA) and Kindlin-2 knockdown (K2 siRNA) BT549 cells (left panel) or MDA-MB-453 cells (right panel) with or without EGF stimulation. Cell nuclei were visualized with DAPI (blue). Arrows mark the nuclear regions. Scale bar: 50 µm. Quantification analysis of the percentage of cells with AR nuclear staining was shown in the lower panel. **p < 0.01, ***p < 0.001 vs. Ctrl siRNA, n = 5 independent experiments for BT549 cells; n = 4 independent experiments for MDA-MB-453 cells. B Luciferase analysis of AR transcriptional activity in control (Ctrl siRNA) and Kindlin-2 knockdown (K2 siRNA) BT549 cells or MDA-MB-453 cells with or without EGF stimulation. Relative luminance was calculated by luminance signal relative to total protein concentration. ***p < 0.001 vs. Ctrl siRNA, n = 9 for BT549 cells; n = 4 for MDA-MB-453 cells. C qPCR analysis of cyclin D1 mRNA expression in control (Ctrl siRNA) and Kindlin-2 knockdown (K2 siRNA) BT549 cells or MDA-MB-453 cells with or without EGF stimulation. *p < 0.05, **p < 0.01, ***p < 0.001 vs. Ctrl siRNA, n = 4. D Immunoblotting analysis of cyclin D1 protein expression in control (Ctrl siRNA) and Kindlin-2 knockdown (K2 siRNA) BT549 cells or MDA-MB-453 cells with or without EGF stimulation (left panel). Quantification of cyclin D1 protein expression was shown at the right panel. **p < 0.01, ***p < 0.001 vs. Ctrl siRNA, n = 4. E Quantification analysis of cell cycle transition in control (Ctrl siRNA) and Kindlin-2 knockdown (K2 siRNA) BT549 cells (left panel) or MDA-MB-453 cells (right panel) with or without EGF stimulation. Cells with 2n signal were in G0/G1 phases; with 4n signal were in G2/M phases; between 2n to 4n signal were in S phase. *p < 0.05, **p < 0.01 vs. Ctrl siRNA, n = 5. F Quantification analysis of cell proliferation assay in the indicated cells, as described in “Materials and Methods”. *p < 0.05, **p < 0.01 vs. Ctrl siRNA, n = 4. G Cell migration was measured by transwell cell migration assay, as described in “Materials and Methods”. Representative images (left panel) and quantification analysis (right panel) were shown. *p < 0.05, ***p < 0.001 vs. Ctrl siRNA, n = 10 for BT549 cells; n = 9 for MDA-MB-453 cells. Original magnification, ×100. K2 Kindlin-2, AR androgen receptor, DAPI 4,6-diamidino-2-phenylindole.

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