Fig. 3.

Transient Lhcgrba expression in teleost germ cells precedes spermatozoon differentiation. (A–E) Bright-field (A) and whole-mount immunodetection (red) of Lhcgrba (B), H3K9ac (C), H4K12ac (D), and α-tubulin (Tuba; E) during spermatid differentiation into spermatozoa after FACS of Senegalese sole SLL cell extracts; the signals are superimposed with the DAPI nuclear counterstain (blue). (F and G) Bright-field (Upper) and double immunostaining of Lhcgrba (green; arrows) and Tuba (red; arrowheads) in zebrafish (F) and gilthead seabream (G) FACS-purified haploid cells; nuclei are counterstained with DAPI (blue). (H) Western blot for Lhcgrba on sole sorted SLL cells. (I) Western blots for Lhcgrba in zebrafish (zf) and seabream (sb) sorted haploid cells. SPD, spermatids; SPDd, differentiating spermatids; SPDc, condensed spermatids; SPZ, spermatozoa. Arrowheads point the spermatozoon flagellum. (Scale bar, 2 µm.)