Skip to main content
. 2022 Apr 12;13:1969. doi: 10.1038/s41467-022-29552-6

Fig. 6. Inactivation of microglial NF-κB rescues tau-induced transcriptomic changes and microglial autophagy deficits while increasing tau inclusions.

Fig. 6

a Representative images of MC1 tau in the hippocampus and cortex of Ikbkb+/+; P301S+ and Ikbkb−/−; P301S+ mice. Scale bar, 250 μm b, c Quantification of the percentage of MC1-occupied area in the hippocampus (b) and cortex (c) of Ikbkb+/+; P301S+ (n = 9) mice and Ikbkb−/−; P301S+ (n = 12) mice; 8 sections/mouse. P-values were calculated using multilevel mixed-effect model with mouse as hierarchical level, ***P < 0.001. d–h Bulk RNA-seq analysis of cortical tissues from Ikbkb+/+, Ikbkb−/−, Ikbkb+/+; P301S+, and Ikbkb−/−; P301S+ mice (n = 4 mice/genotype). DEGs were generated in comparison to Ikbkb+/+ mice. d Heatmap representing DEGs across all genotypes. Normalized and z-scored (color-coded) value of log2PFKM of each gene is shown. Shades of red: upregulation; shades of blue: downregulation. DEGs and biological replicate samples were hierarchically clustered. e Venn diagram comparing the overlap of DEGs in Ikbkb+/+; P301S+ and Ikbkb−/−; P301S+ mice. f Selected IPA canonical pathways identified for 286 DEGs corrected by microglial IKKβ deletion in PS19 mice as in e. P-values were calculated using right-tailed Fisher’s exact test with threshold of significant enrichment as p-value ≤ 0.05 (indicated by a dotted line of −log (p-value) = 1.3). g–k Primary microglia from KFERQ-Dendra mice were incubated with K18/PL tau fibrils with or without TPCA-1. Lysosomal degradation of the CMA reporter was determined by inhibiting lysosomal proteases with NH4Cl and leupeptin (N/L). g Representative images of KFERQ-Dendra microglia co-stained with LAMP1. Left: single channel for Dendra; right: merged channels. Scale bar, 50 μm. Insets: 10 μm. Arrows: Dendra + puncta. UT untreated control. h Quantification of Dendra intensity in microglia incubated with N/L as in g. i Percentage of total LAMP1 puncta positive for Dendra signal in microglia incubated with N/L as in g. j Representative LAMP1 immunostaining in microglia incubated with K18/PL tau fibrils with or without TPCA-1. Scale bar, 50 μm. Insets, 10 μm. k Quantification of average number of LAMP1 positive puncta as in j. h 72; i and k 60 cells from 3 independent experiments, values are mean ± SEM. One-way ANOVA with Tukey post-hoc test. Source data are provided as a Source data file.