NTERA-2 cl. D1 cell were obtained from the American Type Culture Collection (Manassas, VA, USA). Human HMGB4 shRNA was purchased from Fisher Scientific (Pittsburgh, PA, USA). The control shRNA was produced as described50 (link). Cells were infected with lenti-viruses and positive cells were selected after puromycin treatment50 (link). RNA was isolated from the cells, reverse transcribed and analyzed with qBiomarker Validation PCR Array Neuronal Differentiation (Qiagen). The expression level of HMGB4 was analyzed in separate qPCR with specific primers. Protein levels in cells were measured with cell ELISA as described previously53 (link). The following antibodies were used in cell ELISA: mouse anti-β-Actin (clone AC-15; Sigma-Aldrich), mouse anti-GFAP (clone G-A-5; Sigma-Aldrich), mouse anti-NCAM (clone 5B8; Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA, USA), rabbit anti-ASCL1 (Biobryt Ltd., Cambridgeshire, UK), rabbit anti-human HMGB4 (Boster, Pleasanton, CA, USA), rabbit anti-histone H2A acetylated Lys5 and rabbit anti-histone H4 acetylated Lys8 (Cell Signaling Technology, Danvers, MA, USA).
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