Extended Data Fig. 5: Loss of NG2 glia does not alter PrPC expression, prion replication, and prion-induced neuroinflammation.
a-b, Western blots (a) and quantification (b) showing no changes of PrPC level in NG2-glia-depleted (CP673451) Tga20 COCS compared to NG2 glia intact (DMSO) Tga20 COCS. n = 5 samples for DMSO and n = 4 samples for CP673451. Data are presented as mean ± SEM. Unpaired t test (two-sided): P = 0.1231. c-d, Western blots (c) and quantification (d) showing no changes of PK-resistant PrPSc levels in NG2-glia-depleted (CP673451) Tga20 COCS compared to NG2 glia intact (DMSO) Tga20 COCS after prion infection. n = 5 samples for DMSO and n = 4 samples for CP673451. Data are presented as mean ± SEM. Unpaired t test (two-sided): P = 0.3190. e-f, Western blots (e) and quantification (f) showing no changes of PrPC levels in the brains of NG2-glia-depleted (PdgfrαiDTR) mice compared to NG2 glia intact (Ctrl) mice. n = 4 mice for Ctrl and n = 5 mice for PdgfrαiDTR. Data are presented as mean ± SEM. Unpaired t test (two-sided): P = 0.1274. g-h, Western blots (g) and quantification (h) showing no changes of PK-resistant PrPSc levels in the brains of NG2-glia-depleted (PdgfrαiDTR) mice compared to NG2 glia intact (Ctrl) mice after prion infection. NG2 glia depletion was induced at 16 wpi. n = 4 mice for Ctrl and n = 5 mice for PdgfrαiDTR. Data are presented as mean ± SEM. Unpaired t test (two-sided): P = 0.8643. i-j, Cd68 immunofluorescence (i) and quantification (j) showing similar levels of microglia activation in NG2-glia-depleted (CP673451) Tga20 COCS compared to NG2 glia intact (DMSO) Tga20 COCS after prion infection. Nuclei were stained with DAPI (blue). n = 20 slices for NBH + DMSO and Prion+CP673451; n = 17 slices for Prion+DMSO; n = 19 slices for NBH + CP673451. Data are presented as mean ± SEM. One-way ANOVA with Benjamini-Hochberg FDR adjustment for multiple comparisons: P = 0.9795 (NBH + DMSO vs. NBH + CP673451); P < 0.0001 (NBH + DMSO vs. Prion+DMSO); P = 0.5088 (Prion+DMSO vs. Prion+CP673451). k-l, Iba1 and Cd68 immunofluorescence (k) and quantification (l) showing similar levels of microglia activation in the hippocampi of NG2-glia-depleted (PdgfrαiDTR) mice compared to NG2 glia intact (Ctrl) mice after prion infection. NG2 glia depletion was induced at 16 wpi. Nuclei were stained with DAPI (blue). n = 6 mice/group. Data are presented as mean ± SEM. Unpaired t test (two-sided): P = 0.7170 (Iba1); P = 0.8422 (Cd68). m, qRT-PCR results showing minimal changes of proinflammatory factors TNFα, IL1β and IL12β in NG2-glia-depleted (CP673451) Tga20 COCS compared to NG2 glia intact (DMSO) Tga20 COCS after prion infection. n = 6 samples/condition. Data are presented as mean ± SEM. Unpaired t test (two-sided): P = 0.0022 (TNFα); P = 0.3561 (IL1β); P = 0.7870 (IL12β). n, qRT-PCR results showing no changes of proinflammatory factors TNFα, IL1β and IL12β in the brains of NG2-glia-depleted (PdgfrαiDTR) mice compared to NG2 glia intact (Ctrl) mice after prion inoculation. n = 5 mice/group. NG2 glia depletion was induced at 16 wpi. Data are presented as mean ± SEM. Unpaired t test (two-sided): P = 0.1904 (TNFα); P = 0.8378 (IL1β); P = 0.8608 (IL12β).
