Fig. 5

MHCI depletion reduces the activation of primary microglia and motor neuron death in vitro. a Representative SMI-32 immunostaining images of LPS treated and untreated NTG+/+ motor neurons co-cultured with NTG+/+ or NTG−/− microglia. b Cell count analysis showing lower MN loss in LPS treated MNs^NTG+/+- microglia^NTG−/− co-cultures than LPS treated MNs^NTG+/+- microglia^NTG+/+ co-cultures.^*P < 0.05; (LPS_NTG+/+ vs LPS_NTG−/−); ^°°°°P < 0.0001 (LPS_NTG+/+; vs NTG+/+; NTG−/−) by one-way ANOVA with Tukey’s post-analysis. c Representative IBA-1 immunostaining images (red) of LPS treated and untreated microglia cultures from NTG+/+ and NTG−/− mice. d, e Morphometric parameters of IBA-1-positive microglia showing lower activation (mean microglial cell area and circularity) of LPS-treated NTG−/− microglia than LPS-treated NTG+/+ microglia. Data are expressed as mean ± SEM from four independent experiments (at least four stereological 0.6 × 0.6 mm fields analyzed per well). f, g Real-time PCR for Il-23 and Il-1β transcripts from LPS treated and untreated primary microglia cultures from NTG+/+ and NTG−/− mice. Data are normalized to β-actin and expressed as the mean ± SEM fold change ratio between LPS treated NTG+/+ and NTG−/− microglia and relative controls from three independent experiments for each genotype. ^*P < 0.05; ^***P < 0.001 (LPS_NTG+/+ vs LPS_NTG−/−); ^°°°°P < 0.0001 (LPS_NTG+/+; LPS_NTG−/− vs NTG+/+; NTG−/−) by one-way ANOVA with Tukey’s post-analysis. h, i Real-time PCR for Il-23 and Il-1β transcripts in the lumbar spinal cord of G93A+/+ and G93A−/− mice compared to NTG+/+ littermates at 123 and 140 d. Data are normalized to β-actin and expressed as the mean ± SEM fold change ratio between G93A+/+ mice, G93A−/− mice and control mice from four independent experiments for each genotype. ^*P < 0.05; ^**P < 0.01; (G93A+/+ vs G93A−/−); ^°P < 0.05; ^°°°P < 0.001; ^°°°°P < 0.0001 (G93A−/−; G93A+/+ vs NTG) by one-way ANOVA with Tukey’s post-analysis