Journal: International Journal of Molecular Sciences

Article Title: Targeting Cell Death Mechanism Specifically in Triple Negative Breast Cancer Cell Lines

doi: 10.3390/ijms23094784

Figure Lengend Snippet: Validation of the effect of arsenate by qRT-PCR on selected genes related to apoptosis and cell proliferation. Relative gene expression levels are shown for MCL1, BCL2, TGFβ1, CCND1 (Cyclin D1) across cell lines and in arsenate treated and control group (untreated cells). The data were normalized to β-actin and B2M using the ΔΔct method for the HMEC, MCF-7, Hs578T and MDA-MB-231 cell lines compared to the arsenate treated group versus the control group. x is the mean and the line is the median in the boxplots (* p < 0.05). It can be seen that both apoptosis inhibitors (MCL1 and BCL2) are downregulated after arsenate treatment in TNBC cell lines when compared to untreated cells, but not in the normal and DPBC cell lines. Also, survival factor TGFβ1 and cell proliferation indicator CCND1 are downregulated in Hs578T and MDA-MB-231 cell lines compared to their expression in HMEC and MCF-7.

Article Snippet: For immunofluorescence staining, a Human/Mouse BCL2 Antibody (R&D Systems, cat no. AF810-SP, Minneapolis, MN, USA) was used.

Techniques: Biomarker Discovery, Quantitative RT-PCR, Gene Expression, Control, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Targeting Cell Death Mechanism Specifically in Triple Negative Breast Cancer Cell Lines

doi: 10.3390/ijms23094784

Figure Lengend Snippet: ( A ) Microscopy visualization of BCL2 validation at the protein level. Protein expression of BCL2 marked by fluorescently tagged antibodies followed by confocal microscopy evaluation. It can be seen that the expression level of the BCL2 protein is slightly reduced in case of the arsenate treated group compared to control for the case of Hs578T and MDA-MB-231 at 48-h post-treatment, confirming the qRT-PCR and microarray data. ( B ) TGFβ2 and IL6 validation at the protein level. Protein expression of TGFβ2 and IL6 released in cell culture medium 24 h and 48 h for control and arsenate treated cells (HMEC, MCF-7, Hs578T and MDA-MB-231) evaluated by ELISA. x is the mean and median is the middle line in the boxplots. * p < 0.05 two-sided t -test. TGFβ2 is downregulated in protein level after 48 h of arsenate treated cells, but there is no change in the IL6 levels.

Article Snippet: For immunofluorescence staining, a Human/Mouse BCL2 Antibody (R&D Systems, cat no. AF810-SP, Minneapolis, MN, USA) was used.

Techniques: Microscopy, Biomarker Discovery, Expressing, Confocal Microscopy, Control, Quantitative RT-PCR, Microarray, Cell Culture, Enzyme-linked Immunosorbent Assay