aph-2 GSCs have robust expression of Notch downstream target gene sygl-1. a, b) Representative images of distal gonads dissected from Day 1 adult hermaphrodites and Day 5 adult males immunostained for SYGL-1::OLLAS protein. Germ cell nuclei are visualized by DAPI staining. aph-1(tm4743) and aph-2(ik7) animals are M + Z−, and all animals are homozygous for sygl-1(q983[3xOLLAS::sygl-1]). Gonads are oriented with distal end to the left; scale bar is 25 µm. All germ cells in aph-1 gonads are spermatids. c) SYGL-1::OLLAS staining is quantified and plotted according to staining intensity measured in the distal-most five rows of germ cells (n = 20–22 gonads for N2 and aph-2, and n = 15–17 gonads for aph-1), and d) in terms of the number of germ cells that have SYGL-1::OLLAS staining above background (n = 15–16 gonads per genotype for hermaphrodites and n = 19 gonads per genotype for males). Mean (horizontal line) and SD (vertical line) are indicated. P-values are calculated from a Student's 2-tailed t-test, assuming unequal variances.