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. 2021 Aug 19;13(16):4160. doi: 10.3390/cancers13164160

Figure 4.

Figure 4

EFHB and SARAF are required for cell proliferation in breast cancer and pre-neoplastic epithelial cells. (a,b) MCF10A, MCF7, and MDA-MB-231 cells were transfected with shEFHB alone or in combination with EFHB expression plasmid (a) shSARAF alone or in combination with SARAF expression plasmid; (b) or scramble plasmids (shRNAcv as control), as indicated. Forty-eight hours after transfection, cell proliferation was assessed using the BrdU cell proliferation assay kit, as described in Material and Methods, for a further 24, 48, and 72 h. Bar graphs represent cell proliferation 0, 24, 48, and 72 h after the beginning of the experiment, presented as BrdU uptake rate. *** p < 0.001 compared to the corresponding control (cells transfected with shRNAcv). (c) MCF10A, MCF7, and MDA-MB-231 cells were transfected with shEFHB, shSARAF, or scramble plasmids (shRNAcv as control), or were serum-deprived, as indicated. Forty-eight hours later, cell cycle analysis through PI staining and following flow cytometry was performed, as described in “Materials and Methods”. Stacked bars are representative of six independent experiments.