Fig. 5. Restraint stress induces phosphorylation of p38 in testicular germ cells.

a Quantitative real-time PCR analysis of gene expression in different tissues. Averages with s.d. are shown (n = 3 each). b RNA in situ hybridization using dATF-2 mRNA antisense (upper image) and sense (center image) probes. The schematic diagram shows the developmental stages of spermatogenesis (lower diagram). c–e Western blot analysis was performed using samples from w¹¹¹⁸ adult whole-body and testes with anti-P-p38 antibody. c The diagram represents the timing of sampling for the western blots. Samples were prepared at each timepoint (restraint stress for 1, 5, and 10 h and at 12 h after the 10 h restraint stress treatment). Quantitation of the P-p38 from whole-body (d) and testes (e). α-tubulin was used as an internal control. The data represent the mean ± s.d. (n = 3 each) with P values from Student’s unpaired t test: **p < 0.01; N.S., no significant difference. The blotting data were shown in Supplementary Fig. 5b, c. f Immunohistochemical (IHC) staining of P-p38 in testes with/without restraint stress for 10 h. g Representation of average brightness per pixel from the P-p38 signal in testes samples with/without restraint stress. See also Supplementary Figs. 5 and  6.