Figure 8

Wnt3a does not induce the formation of apoptotic nuclei in microglia or compromise cell membrane integrity. Apoptosis was assessed using Hoechst 33342. Microglia on 13 mm coverslips (50000 cells per treatment) were treated with (A) and without (B) Wnt3a (10 nM) for 24 hours then fixed in formaldehyde (4%) and incubated with Hoechst 33342 (17.8 μM). Apoptotic microglia were expressed as a percentage of the total number of cells counted per field (C). Treatments were deemed statistically non-significant (NS) according to a two tailed student’s T-Test. Cell membrane integrity was assessed using fluoroscein diacetate (FDA). Microglia on 13 mm coverslips (50000 cells per treatment) were treated with (D) and without (E) Wnt3a (10 nM) for 24 hours then incubated with fluoroscein diacetate (35 μM: green staining) and Hoechst 33342 (17.8 μM: blue staining) to counter-stain cell nuclei. Fluoroscein diacetate positive cells were expressed as a percentage of the total number of cells counted per field (F). Treatments were deemed statistically non-significant (NS) according to a two tailed student’s T-Test.