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. Author manuscript; available in PMC: 2023 Mar 24.
Published in final edited form as: Free Radic Biol Med. 2018 Dec 13;133:295–309. doi: 10.1016/j.freeradbiomed.2018.12.013

Figure 1. ccRCC cells have increased free iron compared to benign renal cells.

Figure 1.

Free iron levels were estimated with Phen Green staining in four ccRCC cell lines (786–0, A704, 769-P, RCC4) and two primary benign human renal epithelial cell lines (RPTEC, HRCEp) at baseline and after prolonged (120-hour) treatment with and without the iron chelator drug, DFO (500 μM). Intracellular free iron level is inversely proportional to the Phen Green geometric mean fluorescence intensity (gMFI) measured using flow cytometry. (A) Phen Green mean staining levels (gMFI) in untreated ccRCC and benign renal cell lines. (B) Representative flow cytometry histograms of Phen Green stained cells (green) and unstained cells (black) in untreated ccRCC and benign renal cell lines. (C) Phen Green mean staining levels (gMFI) in ccRCC and benign renal cell lines with and without DFO treatment. (D) Representative flow cytometry histograms of Phen Green stained ccRCC and benign renal cell lines with (green) or without (black) DFO treatment. (E) Differences in gMFI levels of DFO treated versus untreated ccRCC cell lines and benign renal cell lines. All data are compiled from at least 3 independent experiments with error bars denoting SEM; *, p<0.05; **, p<0.01 and ***, p<0.001; NS, non-significant. Data in panel (A) were analyzed by one-way ANOVA with Tukey’s post-test, and data in panel (C) were analyzed using two-tailed Student’s t-test comparing treated and untreated conditions for each cell line. For panels (A) and (E), the top and bottom asterisk labels denote significance compared to HRCEp and RPTEC, respectively.