Fig. 3

AST factor-mediated globin induction confers anoikis resistance. A Measurement of cell detachment-induced ROS production in HEK293A-4AST (right) versus trypsinized cells (left). Green, ROS. Scale bar, 50 μm. B Heatmap of genes involved in ROS defense mechanisms in HEK293A-mock and 4AST cells. C Quantitative real-time PCR analysis of the HBA1/2 transcript levels in 4AST cells. D Analysis of H3K27ac ChIP-seq profiles at HBA1 and HBA2 loci in HEK293A-mock and 4AST cells. E Representative fluorescence images of ROS accumulation in HEK293A-4AST cells treated with scramble or HBA1/2 siRNAs with or without N-acetylcysteine (NAC). F Measurement of cell viability of HEK293A-4AST cells upon siRNA-mediated HBA1/2 depletion. G Fluorescence images of ROS accumulation in HEK293A-4AST cells by hemin treatment. Scale bar, 50 μm. H Measurement of cell viability in HEK293A-4AST cells upon hemin treatment. n = 3. I Schematic of mechanistic insights into AST factor-mediated reprogramming of anchorage dependency. For (C and F) mean values are shown with error bars representing the s.d. of n = 3 independent replicates. *p < 0.05