Extended data 6. Translation blockade does not affect cleaved caspase 3 staining in acute slice setting.

a) Whole mount immunostains were prepared from acute slices as in Figure 4d–i except immunostained with TMEM119, cleaved Caspase 3, and DAPI. Two slices were treated with 3% ethanol (30 minute treatment) as a positive control. A TMEM119+ microglial soma (white arrow) that has a PeMP encircling and forming a phagocytic cup around a cleaved caspase-3 positive apoptotic non-microglial cell (red arrow) within the ethanol-treated group. Images representative of 2 independent experiments. b-c, No significant differences were detected between anisomycin and vehicle-treated slices in measurements of cleaved caspase-3 postive area (b, p = 0.82 by unpaired two-tailed t-test), or cleaved caspase-3 colocalized with microglial marker, TMEM119 (c, p = 0.26 by unpaired two-tailed t-test). N = 9 vehicle, n = 7 anisomycin, and n=2 ethanol treated slices derived from n = 2 mice processed and stained on separate days. Data are shown as mean ± s.e.m.