Figure 6. Recombinant celsr2 EC4–8 protein induced germ cell detachment from Sertoli cells in vitro. Immunohistochemical localization of celsr2 in Sertoli cell–germ cell co-cultures showed celsr2 punctate staining at areas in which clusters of spermatocytes adhered to the Sertoli cell monolayer (A; arrow). Preabsorbed negative control is shown in B. Bar = 5 µm. (C) Sertoli cell–germ cell co-cultures were exposed to vehicle (negative control), recombinant celsr2 EC4–8 protein, recombinant celsr3 EC4–8 protein, or 100 µM mono-(2-ethylhexyl)phthalate (MEHP)(positive control). Negative control germ cell detachment at each time point was set to 1, and detachment in treatment groups were expressed as a ratio to negative control. Columns with the same letter designation were not significantly different from each other, determined via a 1-way analysis of variance (ANOVA) followed by Tukey's multiple comparisons test.