Fig. 4

G-CSF exacerbates spermatogenic loss before busulfan treatment but enhances recovery after busulfan treatment. Results are from animals in Experiment 3. a Testis weights (mean ± standard error). Significant differences were determined by a Student’s t-test and are noted by asterisks above noted bars (*p ≤0.006). b Cauda epididymal sperm counts (swim-up, mean ± standard error; **p ≤0.002). Micrographs of individual seminiferous tubule cross-sections demonstrating complete spermatogenesis (c-e) stained with H&E, or with antibodies against (f-h) γH2A.X (spermatocyte marker) and (i-k) lectin peanut agglutinin (PNA; spermatid marker) are shown from representative sections of testes from (c, f, i) Control, (d, g, j) Busulfan only group and (e, h, k) Busulfan + G-CSF group. Scale bars = 20 μm. l Stacked bars show the percentage of all seminiferous tubule cross-sections counted from all animals in each group which exhibit differing degrees of spermatogenesis: complete spermatogenesis (complete), up to round spermatid spermatids, up to 1° spermatocytes, or containing no spermatogenesis (empty or Sertoli cell-only), ***p ≤0.086). The number of seminiferous tubule cross-sections evaluated per animal is shown in Additional file 1: Table S1. m The proportion of seminiferous tubule cross-sections containing complete spermatogenesis in each individual animal (number indicated below x-axis), grouped by treatment group