Figure 7: Mgst2 deficiency attenuates ER stress-triggered oxidative stress, DNA damage and apoptosis.

(a) Agarose gel electrophoresis of PCR products obtained by amplification of DNA isolated from tail ends of WT and homozygous Mgst2-deficient (KO) mice. DNA of heterozygous ES cells (ES) and negative PCR control (Neg.) are also shown. The 426-bp band corresponds to WT DNA sequence from the Mgst2 gene. The 805-bp band corresponds to the mutated allele. (b,c) Immunostain of LTC₄ in WT and Mgst2-deficient MEFs at passage 2 following treatment with BfA. Bar, 20 μm. n=3, ***P<0.0001. (d) Immunostain of NOX4 in WT and Mgst2-deficient MEFs at passage 2, treated with vehicle or Tm (4 μg ml⁻¹, 24 h). Bar, 20 μm. This image is a representative of five replicates. (e,f) ROS detection using DCFH-DA in primary WT and Mgst2-deficient (KO) MEFs, treated with vehicle or BfA (0.25 μg ml⁻¹). Bar, 20 μm. n=4, ***P<0.001. (g,h) Immunostain of 8-OHdG in primary WT and Mgst2-deficient MEFs, treated with vehicle or BfA. Bar, 20 μm. n=3, ***P<0.001. (i) Immunoblot of the indicated proteins in WT and Mgst2-deficient primary MEFs, treated with Tm (3 μg ml⁻¹). (j) Cultures of WT and Mgst2-deficient primary MEFs, treated with vehicle or Tm (2 μg ml⁻¹) and then stained with crystal violet. Bar, 50 μm. (k) Relative viability as determined by neutral red staining of MEFs, treated as in j. n=3, **P=0.005. Images a and i are representatives of at least three replicates. Values in c,f,h and k represent the mean±s.d. Statistical significance was determined using one-way ANOVA.