Fig. 1

Histone H4 acetylation mutants. a Western blot analysis using total protein from the transfectants grown at 2.5 and 10 µg/ml blasticidin to check the presence of HA-tagged H4 proteins using antibodies against HA. Histone antibodies were used as positive control to confirm the expression of endogenous histones and actin was used as a loading control. b Western blot analysis carried out from cytoplasmic, low salt nuclear, high salt nuclear and total SDS fractions to confirm nuclear localization of HA-tagged H4 proteins in the transfectants. c Microarray expression analysis for transfectants grown at 10 µg/ml blasticidin. Heat maps represent the differentially expressed genes in the transfectants at each stage (Student’s t test, P < 0.05). Data shown is log₂ ratio of average of biological triplicates, normalized to H4-HA transfectant. d Functional pathways significantly up or downregulated in the differentially expressed gene sets (data from c) of the transfectants. The scale shows the fold enrichment of number of genes in the specific pathways [based on hypergeometric test (P < 0.05)]. e Effect of H4 mutations on multiplication. Cells were grown in 2.5 or 10 µg/ml blasticidin and FACS using Hoechst stain was carried out to count the number of new rings at each invasion cycle for 6 cycles. f The scatter plot of merozoite number in the segmented schizonts in H4ac4R, H4K8R and H4-HA transfectants determined by Giemsa stain light microscopy