Extended Data Fig. 7: The rescue of neuronal vulnerability by GSK is more effective overall in the surface of C9 CO slices.

a, Representative low magnification immunofluorescence images of γ-H2AX⁺ foci in CTIP2⁺ deep layer neuronal (DLN) and SOX9⁺ astroglial nuclei in C9 CO slices immersed in media and treated with GSK or vehicle in comparison to untreated controls for experiments quantified in b (8 independent untreated and 6 treated COs). Arrowheads illustrate nuclei (dashed circles defined by DAPI staining) with γ-H2AX⁺ foci (green). b, For C9-L1 CO slices (240DIV), graphs represent foci/cell ± s.d.; n = 105, 110 and 32, 32 CTIP2⁺ DLNs and 57, 23 and 25, 14 SOX9⁺ astroglia in surface and middle sections for C9 and GSK-treated C9 independent CO slice-pairs, respectively. For C9-L2 COs (200DIV), graphs represent foci/cell ± s.d.; n = 640, 534 and 404, 286 CTIP2⁺ DLNs and 631, 558 and 411, 274 SOX9⁺ astroglia in surface and middle sections for untreated and GSK-treated independent C9 CO slice-pairs, respectively; Two-sided Mann Whitney test for the GSK effect. c, Quantification of pyknosis in CTIP2⁺ DLN and SOX9⁺ astroglial nuclei in C9 CO slices treated with GSK or vehicle and in untreated controls. Data represent the proportion of pyknotic nuclei with residual CTIP2- or SOX9-immunoreactvity and expressed as mean ± s.d. for surface or middle sections; n = 3 (2 C9-L1/1 C9-L2) untreated and 3 GSK-treated independent C9 CO slice-pairs at 240 DIV (upper panel) or 3 C9-L2 untreated and 3 GSK-treated independent CO slice-pairs at 200DIV (lower panel) per group; Two-tailed unpaired t-test for the GSK effect. Scale bars=10 μm. See Supplementary Table 3 for detailed statistics.