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. 2024 Jan 31;10(5):eadj9479. doi: 10.1126/sciadv.adj9479

Fig. 1. Folate depletion induces differentiation in erythroid cancer cell lines.

Fig. 1.

(A) Experimental timeline (bottom) and proliferation (top) of K562 cultured for 6 days in folate-free RPMI supplemented with 2000 or 100 nM FA. (B) Simplified schematic of de novo purine synthesis. (C) Volcano plot depicting day 6 metabolic changes between K562 cells cultured in 2000 and 100 nM FA. PRPP, phosphoribosyl diphosphate; dTMP, deoxythymidine monophosphate. (D) Schematic of the human heme biosynthesis pathway. (E) Reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis of heme biosynthesis genes in 2000 nM or 100 nM FA. (F) RT-qPCR analysis of ALAS2 in the indicated cell lines following 6 days culture in 2000 or 100 nM FA. (G) Western blot analysis of hemoglobin (Hb) A1 and fetal expression in K562 over 8 days in 100 nM FA. (H) Cell surface GlyA levels in K562 measured by flow cytometry. MFI, mean fluorescence intensity. (I) Western blot analysis of hemoglobin A1 in MEL. (J) Cell surface Ter119 levels in MEL as measured by flow cytometry. (K) Western blot analysis of hemoglobin A1 in HEL. (L) Cell surface GlyA levels in HEL as measured by flow cytometry. All data shown are means (±SD) of three biological replicates except (E) n = 3 to 4 and (F) n = 2 to 3 (K562 and MEL).