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. 2020 Feb 1;8(4):493–508. doi: 10.1016/j.gendis.2020.01.015

Figure 6.

Figure 6

ACLY and Cyclin D1 are CD49b targets enhancing self-renewal and cell cycle. (A) Immunoblot of downregulated ACLY proteins in MDA-MB-231 cells transfected with siACLY and siITGA2 in comparison to siRNA control and siOCT4. (B) Reduced cellular acetyl-coA levels in the ITGA2-knock down MDA-MB-231 cells, as measured by LC/MS (t-test P = 0.001, n = 3 biological replicates). Error bars represent S.D. values. (C) Compromised mammosphere formation in ACLY KD cells, shown representative images (right panels) and quantified histograms (left panel) (t-test P < 0.05, n = 3 biological replicates). Error bars represent S.D. values. Scale bars = 30 μm. (D) Immunoblot of downregulated cyclin D1 proteins in MDA-MB-231 cells transfected with siITGA2 compared to control. (E) siITGA2-induced G1 arrest and altered G2/M phase were rescued by CCND1 overexpression in MDA-MB-231 cells. *P < 0.05, **P < 0.01 (t-test), n = 3 biological replicates. Error bars represent S.D. values. (F) Representative images (top panels) and qualification (bottom panel) of mammosphere formation of MDA-MB-231 cancer cells upon siITGA2 mediated KD. ** t-test P < 0.01, n = 3 biological replicates. Error bars represent S.D. values. Scale bars = 50 μm. (G) Bioluminescence images of siControl, siACLY, siCCND1-transfected MDA-MB-231 tumor cells upon tail vein infusion (n = 4 mice each group). (H) Fold change of lung metastasis signals (total photons, p/s), normalized to day 0 (D0). t-test P = 0.003 (**), 2e-5 (****), and 7e-5 (****), for D1, D2, and D3 comparisons between the control and siCCND1 groups, respectively. Error bars represent S.D. values.