Fig. 3. CAR-1 localization in the germline. Extruded gonads from one-day-old hermaphrodites were analyzed by immunostaining for CAR-1 (A) and DAPI staining for DNA (B). The distal region is to the bottom left and the proximal region at the top right. CAR-1 levels are low in the distal mitotic zone but increase as the germ cells enter meiosis (see detail). (C-J) CAR-1 localization during the pachytene stage. Extruded gonads were stained for CAR-1, CGH-1 and PGL-1. (C) Perinuclear localization of CAR-1; (D) merge of CAR-1 with CGH-1 and DAPI staining. (E-G) Merged CAR-1 (green) and CGH-1 (red) staining (E); merged CAR-1 (green) and PGL-1 (red) staining (F); PGL-1 staining alone (G); images are from the area surrounding the nucleus that is indicated in D. The extent of overlap between CGH-1, CAR-1 and PGL-1 staining was reproducibly comparable to that shown, but among individual foci the degree of overlap and the relative orientation of CGH-1- and CAR-1-stained foci varied. (H-J) A cross-section through the germline core, shown to highlight cytoplasmic CAR-1 (H) and CGH-1 (I) foci; a merged image with DAPI staining is shown in J. (K-N) Specific mislocalization of CAR-1 in the cgh-1(ok492) gonad, revealed by antibody staining. In the mitotic region (K), CAR-1 is present at low levels in perinuclear foci that colocalize with PGL-1 (L), as in wild type (not shown), but its localization becomes dramatically altered within the transition zone (TZ). (K-N) DAPI staining; (K-M) CAR-1 staining (green); (L) merge, including PGL-1 (red), which corresponds to the boxed region in K. Within the pachytene region of cgh-1(ok492) germlines, CAR-1 localization is highly abnormal (M), but the levels and localization of PGL-1 antibody staining are not detectably altered (N). (C-L,N) Single plane confocal images; (M) a confocal z-series projection.