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. 2013 Oct 20;19(12):1305–1320. doi: 10.1089/ars.2012.5021

FIG. 4.

FIG. 4.

High proliferative cancer cells show higher GSH levels and glutathionylated H3 than normal fibroblasts. Gene transcription of proliferation related genes is affected when GSH is depleted. (A) Immunoblots of histone extracts from HeLa and MDA-MB-468 tumor cell lines and normal human fibroblasts (HF1 and HF2) incubated with anti-GSH and anti-H3 antibodies under nonreducing conditions. (B) GSH cellular levels in tumor and normal cells determined by GSH S-transferase assay. (C) GSH levels in NIH3T3 cells at 24 h after GSH depletion by DEM treatment by GSH S-transferase assay. (D) DNA synthesis rates for control (nontreated) and DEM treated NIH3T3 cells at 24 h of cell culture. (E) Immunoblot analysis of glutathionylated histones showing the band corresponding to H3 (top) and the replication-dependent H3.1 after GSH depletion by DEM in NIH3T3 at 24 h of cell culture. Equal loading was controlled using anti-H3 antibody in the immunoblot (bottom). (F) Relative expression levels of HIST1H3A, PCNA, and Id2 genes by qRT-PCR after GSH depletion using DEM (GAPDH expression was used for normalization). The statistical significance is expressed as: *p<0.05, #p<0.05, &p<0.05 between compared groups. DEM, diethyl maleate.