Figure 2.

NECs undergo replication stress upon in vivo defective licensing

(A) Coronal brain sections of mouse embryos at E9.5 immunostained with anti-γΗ2ΑΧ (red) and anti-Sox2 (green). Cortical walls are restricted by the dotted lines. White boxes mark the higher-magnification images shown on the right. The demarcated cells represent positive cells. Note the γΗ2ΑΧ foci that are formed, representative of DNA damage.

(B) The graph depicts the γH2AX⁺ cells per 200-μm-wide area. Mean ± SEM (n = 3/genotype).

(C) Expression of 53BP1 (green) in the cortical wall of embryos at E10.5. The arrows indicate positive cells demarcated in higher-magnification images.

(D) The percentage of 53BP1-positive cells with >4 discernible foci per nucleus is presented in the bar graph. Mean ± SEM (n = 3/genotype).

(E) pATR (green) in brain sections of E9.5 embryos. The boxes indicate higher magnifications shown on the right.

(F) The graph shows the cells that express pATR per 200-μm-wide column. Mean ± SEM (n = 3/genotype).

(G) DNA fibers were spread from the cortexes of E10.5 embryos. The lengths of individual tracks were quantified and are presented in the graph (n = 46 control, n = 39 Gmn;Foxg1; data derived from two independent litters).

(H) The lengths of the green tracks in bidirectional forks were quantified in DNA fibers and the ratio of the sister forks is presented in the graph (n = 14 control, n = 19 Gmn;Foxg1; data derived from two independent litters). DNA is stained with Hoechst (blue). Scale bars: 10 μm. ^∗p ≤ 0.05, ^∗∗p ≤ 0.01; t test.