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. 2023 Oct 4;4(10):101211. doi: 10.1016/j.xcrm.2023.101211

Figure 2.

Figure 2

High A3B levels cause male-specific infertility

(A) Progeny numbers and A3B status for the indicated crosses (n = 3 litters per cross). In parental animals with A3B, the indicated A3B minigene (R26 or CAG) is heterozygous in combination with a WT Rosa26 locus (data not shown). The A3B status of progeny is dictated by the parental cross.

(B) Images of a representative testicle and epididymis from WT and CAG-A3B males.

(C and D) H&E-stained sections of WT (top) and CAG-A3B (bottom) testicle and epididymis, respectively.

(E and F) Anti-A3B IHC staining of the seminiferous tubule and epididymal lumen from WT and CAG-A3B males, respectively.

(G and H) Representative images and quantification of spermatozoa from WT and CAG-A3B males, stained with eosin-nigrosin to distinguish live (white) and dead (pink) cells, respectively (mean ± SD of n = 200 sperm from 3 independent males; unpaired t test p value indicated).

(I) Images of zygotes 7 h postfertilization of a WT ovum with spermatozoa from the indicated male genotypes. Arrows point to pronuclei, which indicate successful fertilization.

(J) Proportion of embryos at the indicated developmental stage 48 h postfertilization in vitro (n > 50 zygotes analyzed per condition).

(K) Images of developing embryos 96 h postfertilization in vitro.

(L) Proportion of embryos at the indicated developmental stage 96 h postfertilization in vitro (n > 50 zygotes analyzed per condition; continuation of experiment reported in J).