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. 2021 Jan 7;10(1):97. doi: 10.3390/cells10010097

Figure 2.

Figure 2

Lack of Fer and FerT renders metastatic lung cancer cells susceptible to metformin treatment. (A) Fer- and FerT-deficient HΔFer/FerT cells were generated using the CRISPR-Cas9 system. Western blot analysis of lysates from parental H1299 cells expressing both Fer and FerT proteins, and six derived HΔFer/FerT knock-out clones (#3, #12, #16, #35, #42, and #45) are shown. Actin served as a loading control. (B) Parental H1299 and HΔFer/FerT cells were either left untreated (NT) or treated with (C) 5-mM metformin or (D) 10-mM metformin for 24 h. Cells were stained with annexin V/PI followed by flow-cytometry analysis. Annexin V staining (Q3) indicates apoptotic cell death. PI staining (Q1) indicates necrotic cell death. Annexin V/PI double staining (Q2) indicates late apoptotic/early necrotic cell death. Each experiment represents one out of three independent experiments that gave similar results. ** denotes p < 0.01.