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. Author manuscript; available in PMC: 2023 Aug 4.
Published in final edited form as: Mol Cell. 2022 Jun 16;82(15):2832–2843.e7. doi: 10.1016/j.molcel.2022.05.024

Figure 3. SLCO2B1 expression is sufficient to promote iron availability and sustain cell proliferation under iron restriction.

Figure 3.

(A) Fold change in the number (log2) of vector, SLCO2A1 and SLCO2B1-expressing cells after 5 days in the presence of indicated DFO concentrations (left). Representative bright-field micrographs of Jurkat cells after 5-day treatment with 3 μM DFO (right). Scale bar is 50 μm.

(B) Fold change in the number (log2) of vector, SLCO2A1 and SLCO2B1-expressing cells after 5 days in the presence of indicated BafA1 concentrations (left) and representative bright-field micrographs of Jurkat cells after 5-day treatment with 4 nM BafA1 (right). Scale bar is 50 μm.

(C-E) Fold change in the number (log2) of vector and SLCO2B1-expressing cells after 5 days in the presence of indicated (C) DFP, (D) NH4Cl and (E) ConA concentrations

(F) Schematic (top) showing IRP2-mediated regulation of ferritin heavy chain (FTH1). Increased iron levels inhibit IRP2, a negative regulator of FTH1 expression. Immunoblot analysis of FTH1 and SLCO2B1 in indicated human and mouse cell lines expressing vector or SLCO2B1 cDNA (bottom). β-actin was used as loading control.

(G) Immunoblot analysis of IRP2 and FTH1 in Jurkat and 293T cells expressing vector or SLCO2B1 cDNA, in the absence or presence of iron depleting agents BafA1 (3 nM in Jurkat; 10 nM in 293T), NH4Cl (1 mM in Jurkat; 5 mM in 293T) or DFO (3 μM in Jurkat; 100 μM in 293T).

(H) Abundance of indicated metals in Jurkat cells expressing vector or SLCO2B1 cDNA. Results were normalized to total amount of protein and reported as ‘pg metal per μg protein’.

(I) Representative bright-field micrographs of TFRC knockout Jurkat cells expressing vector, SLCO2A1 or SLCO2B1 cDNA. Scale bar is 50 μm.

(J) Fold change in the number (log2) of TFRC knockout Jurkat cells expressing a control vector, SLCO2A1 or SLCO2B1 cDNA without (top) or with 0.01 mg/ml ammonium ferric citrate (FAC) supplementation (bottom) after 3 days. (Mean ± SEM, n= 3, Student’s t-test, 95% confidence interval). See also Figure S3