Fig 2: Effect of the BGP-15 treatment on the regulation of mitochondrial biogenesis. Representative Western blot analysis of PGC-1α, VDAC, CREB, AMPK, and phosphorylation of CREB and AMPK. Densitometric evaluation is also shown. GAPDH was used as a loading control. WKY: age-matched normotensive Wistar-Kyoto rats, n = 7; SHR-Baseline: 15-month-old spontaneously hypertensive rats, n = 7; SHR-C: nontreated spontaneously hypertensive rats, n = 7; SHR-B: spontaneously hypertensive rats receiving BGP-15 for 18 weeks, n = 7. Values are mean ± SEM. ∗p < 0.05 vs. WKY, ∗∗p < 0.01 vs. WKY, ##p < 0.01 vs. SHR-Baseline, §p < 0.05 vs. SHR-C, §§p < 0.01 vs. SHR-C.

Fig 3: Western blot analysis of CREB and p-CREB. a The representative immunoblot made from hippocampal tissue of rats. b The quantification of CREB/β-actin ratio levels. c The quantification of p-CREB/β-actin ratio levels. ★ P < 0.05, ★★ P < 0.01 vs. control group; ▲ P < 0.05 vs. CUMS group; ◆ P < 0.05 vs. Acu group; ■ P < 0.05, ■■ P < 0.01 vs. FLX group. (mean ± SD, n = 6)

Fig 4: Quantification of the expression of genes and proteins involved in synaptic plasticity. Genes significantly changed in expression from hippocampus and cortex at doses of 0.5 Gy and 1.0 Gy using RT2 Profiler PCR Arrays are shown in A. The table shows the genes which are significantly up-regulated or down-regulated with the fold-changes in brackets; *p < 0.05; **p < 0.01; ***p < 0.001 (unpaired Student’s t-test, n = 3) after normalisation to the median of 84 target genes. Immunoblots and quantification of Arc, c-Fos, total CREB and phosphorylated CREB in hippocampus and cortex of sham-irradiated and 1.0 Gy irradiated mice 7 months post irradiation (B and C). The columns represent the fold-changes with standard errors of the mean (SEM) from three biological replicates. The visualisation of protein bands shows the representative change using three biological replicates. *p < 0.05; **p < 0.01; ***p < 0.001 (unpaired Student’s t-test). Normalisation was performed against endogenous GAPDH; H: Hippocampus, C: Cortex.

Fig 5: nsPEFs activated JNK, CREB, and STAT3 signaling pathways. a Western blot analysis for JNK, p-JNK with inhibitors for phosphorylation of JNK (BI-78D3), and CREB (BAPTA-AM). b Phosphorylation ratio for JNK with inhibitors, n = 5. c Western blot analysis for CREB, p-CREB with inhibitors for phosphorylation of JNK (BI-78D3), and CREB (BAPTA-AM). d Phosphorylation ratio for CREB with inhibitors, n = 5. e Western blot analysis for STAT3, p-STAT3 with inhibitors for phosphorylation of JNK (BI-78D3), CREB (BAPTA-AM), and STAT3 (Stattic). f Phosphorylation ratio for STAT3 with inhibitors, n = 5. Statistical significance in mean values was marked with different letters. For all variables with the same letter (a, b, c, d, or e), the difference between the groups is not statistically significant. For variables with a different letter, the difference between the groups is statistically significant (P < 0.05)