Figure 4. Transcriptional upregulation of sex chromosome-linked genes in pachytene spermatocytes from rapamycin-treated mice. (a) Average expression levels (rapamycin versus control) from chromosomes. RNA-seq was performed on RNA from pachytene spermatocytes isolated from adult control (n=7) and rapamycin-treated (n=15) mice using the STA-PUT method. (b) Quantitative RT-PCR analysis of X and Y-linked genes in pooled samples of pachytene spermatocytes used in (a). Levels of Arbp mRNA were used as loading control. *P<0.05, ** P<0.01. Student’s t test. (c-d) Mean expression levels of autosomal (A), X-linked (X) and Y-linked (Y) genes in control pachytene spermatocytes (c) and round spermatids (d). (e) Average expression levels (rapamycin versus control) for each chromosome in round spermatids isolated from adult mice (control, n=7; rapamycin, n=15). *P<0.05, ** P<0.01.