Immunoblotting of proteins were performed by following standard protocol. Briefly, total protein from HUDEP2 cells at day 5 of differentiation or primary cells at day 13 of differentiation were extracted with RIPA buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS, 1 mM EDTA)58 (link). Equivalent amounts of protein (10-20 µg) were loaded onto 4-12% or 10% Bis-Tris protein gels (Invitrogen) and separated in 1× MES-SDS running buffer (Invitrogen, NP0002). Separated proteins were transferred onto PVDF membrane and incubated with primary antibodies overnight. Primary antibodies used were anti-NFIA (Active Motif, #39397), 1:5,000; anti-NFIX (clone 3D2, Sigma Aldrich, SAB1401263), 1:500; anti-NFIC (Cell Signaling, #11911), 1:1,000; anti-BCL11A (Abcam, ab19487), 1:1,000; anti-LRF (Thermo Fisher Scientific, #14-3309-82), 1:2,000; anti-hemoglobin γ (Santa Cruz, #21756), 1:1,000; anti-β-Actin (Santa Cruz, #47778), 1:1,000; anti-HA (Cell Signaling, #3724), 1:1,000.