Journal: Journal of Neuroinflammation
Article Title: Natural killer cells aggravate neuroinflammation through microglial type I interferon pathway activation after traumatic brain injury
doi: 10.1186/s12974-026-03715-4
Figure Lengend Snippet: NK cell depletion alleviated the activation of type I interferon signaling pathway in microglia. A - C Transcriptional analysis of M1-markers, IFN pathway markers and M1-markers of isolated microglia. n =4 animals per group. D , E Representative western blotting bands and quantification of STAT1, IFIT3,IRF7 in the peri-contusional cortices from mice subjected to sham, TBI, TBI+ NK cell depletion ( F ) Representative images of immunofluorescent staining for CD16/32 (green), Iba-1 (red), and DAPI (blue) in the ipsilateral cortex at 3 days post-TBI. G Representative images of immunofluorescent staining for CD206 (green), Iba-1 (red), and DAPI (blue) in the ipsilateral cortex at 3 days post-TBI. H Representative FACS plots showing gating strategy we used in flow cytometric analysis. CD11b+ CD45 int (microglia) were gated for further analysis. H Quantification of CD16/32+ and CD206+ subsets of microglia in Sham, TBI and TBI+NK depletion. Data are presented as mean ± SEM, statistical analysis with one-way ANOVA followed by Tukey post-hoc test in A - C , E , J . *comparison of TBI versus sham, #comparison of TBI versus TBI+NK depletion. ns, not signifcant; #, p <0.05, ##, p <0.01, ###, p <0.001, *, p <0.05; **, p <0.01, ***, p <0.001. Scale bars: 50 µm
Article Snippet: Then, the membrane was incubated with primary antibodies against anti-STAT1(1:1000; ab288157; Abcam), anti-IRF7(1:1000; sc-74471; Santa Cruz), anti-IFIT3(1:1000; sc-393396; Santa Cruz), anti-GRP78 (1:1000; sc-13539; Santa Cruz), anti-Phospho-eIF2α-S51 (1:1000; 87179-2-RR; Proteintech), anti-eIF2α (1:1000; 11170-1-AP; Proteintech), anti-Phospho-PERK (Thr980) (1:1000; 16F8; Cell Signaling Technology), Anti-PERK (1:1000; C33E10; Cell Signaling Technology), anti-ATF4 (1:1000; D4B8; Cell Signaling Technology), anti-GAPDH (1:1000; ABclonal), anti-GAPDH (1:1000; ABclonal) at 4 °C overnight.
Techniques: Activation Assay, Isolation, Western Blot, Staining, Comparison
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