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Fig. 2 | BMC Cancer

Fig. 2

From: Dishevelled 2 regulates cancer cell proliferation and T cell mediated immunity in HER2-positive breast cancer

Fig. 2

Nuclear DVL2 binds to promoter regions and regulates transcription of infiltrating T-cell genes in HER2+ breast cancer cell lines. A Immunofluorescence staining was performed to analyze DVL2 proteins localization in SKBR3 (ER−/PR−/HER2+) breast cancer cells. The cells were probed with DVL2 (red). The nucleus was stained with DAPI (blue) and the actin filaments (green) were stained with Phalloidin. B Whole (WCE), cytoplasmic (Cyto) and nuclear (Nucl) extracts from SKBR3 breast cancer cells were analyzed using western blots. The blots were probed with DVL2 antibody. Lamin was used as a control for nuclear extract and Tubulin was used as a control for cytosolic proteins. C ChIP PCR was performed for different immune-modulatory genes for Input, IgG, DVL2 and DVL3 in SKBR3 breast cancer cells. D RT-qPCR based mRNA expression analyses of DVL2, HLA-C, STAT1, TGFB1, STAT6 in SKBR3 cells stably expressing NTC vs. shDVL2. Transcript levels were normalized using GAPDH, B2M and GUSB as housekeeping control. All experiments were performed in triplicates and Welch’s independent T-test were performed between NTC and shDVL2 where *p < 0.05 and #p < 0.1. E Immunofluorescence staining was performed to analyze DVL2 proteins localization in BT474 (ER+/PR+/HER2+) breast cancer cells. The cells were probed with DVL2 (red). The nucleus was stained with DAPI (blue) and the actin filaments (green) were stained with Phalloidin. F Whole (WCE), cytoplasmic (Cyto) and nuclear (Nucl) from BT474 breast cancer cells were analyzed using western blots. The blots were probed with DVL2 antibody. Lamin was used as a control for nuclear extract and Tubulin was used as a control for cytosolic proteins. G ChIP PCR was performed for different immune-modulatory genes for Input, IgG, DVL2 and DVL3 in BT474 breast cancer cells. H RT-qPCR based mRNA expression analyses of DVL2, HLA-C, STAT1, TGFB1, STAT6 in BT474 cells stably expressing NTC vs. shDVL2. Transcript levels were normalized using GAPDH, B2M and GUSB as housekeeping control. All experiments were performed in triplicates and Welch’s independent T-test were performed between NTC and shDVL2 where *p < 0.05 and #p < 0.1

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