Figure 7

Evaluation of neuroinflammation, astrogliosis and oxidative stress. Immunostaining for CD11b showing significantly increased expression in dentate gyrus (DG), molecular layer (ML) and hilus (HL) after irradiation with 1.0 Gy compared to sham-irradiated controls (A and B) is shown. Gene expression analysis/immunoblots of TNFα, phospho-IGF1Rβ/INSRβ within the hippocampus [H] is shown (C and D). Figure E and F show immunostaining for GFAP with a significant increase in the number of astrocytes in the hilus after irradiation with 0.1 Gy, 0.5 Gy and 1.0 Gy compared to sham-irradiated controls. Immunoblot of total malondialdehyde (MDA)-tagged proteins within the hippocampus [H] is shown (G and H). The visualisation of protein bands shows the representative change from the biological replicates; MDA content quantification was performed with 5 bands in the range of 35 kDa to 55 kDa after a total lane normalisation. All data are reported as mean/fold-change ± SEM (n = 5 for immunohistochemistry; n = 4 for immunoblotting against MDA content; n = 3 for gene expression and other immunoblots); *p < 0.05; **p < 0.01; ***p < 0.001 (unpaired Student’s t-test). Immunoblot of total malondialdehyde (MDA)-tagged proteins within the cortex [C] exposed to sham-irradiation and 1.0 Gy (I) and quantification of six representative bands in the range of 25 – 70 kDa (J) are shown. Statistical analysis was performed with unpaired Student’s t-test. Data are reported as fold-change ± SEM (n = 4). The quantification of the protein bands was done using four biological replicates.