Fig. 6: 3xDR microglia exhibit an aged-like phenotype.
From: Transcriptional and epigenetic decoding of the microglial aging process
a, Scheme of 3xDR and timepoints for experiments. b, 3xDR shortens the microglial telomere. n = 7 mice for each group. Two-tailed independent t-test. c, 3xDR microglia (1,313 cells) exhibited transcriptional characteristics distinct from control mice (2,180 cells). n = 5 mice for each group. d, 3xDR microglia display higher P-ADEM and lower N-ADEM gene signature scores. e, 3xDR versus control microglia FC (qPCR) exhibited a similar trend to LPS challenge as aged versus young microglia FC (bulk RNA-seq). n = 4 (3xDR and control) and 3 (3-month-old and 24-month-old; data from Fig. 4) mice. Linear regression. f, tSNE plots show the expression levels of microglial activation-associated and homeostasis-associated genes. g, 3xDR microglia exhibit a phenotype resembling that of aged microglia. h, 3xDR and 24-month-old microglia exhibit higher pseudotime value than 3-month-old and 14-month-old microglia. One-way ANOVA with Bonferroni’s post hoc test. n = 5 mice for each group. In total, 4,207 young, 3,272 middled-aged, 2,497 aged and 1,313 3xDR microglia (g,h). i, Only five genes are overlapping between 3xDR DEGs and ARM gene set. j–l, 3xDR microglia exhibit a distinct morphology to control microglia. 3xDR microglia display a larger cell body than control microglia. n = 8 mice for control and 9 mice for 3xDR, 100 cells from cortex and hippocampus for each group. Each dot represents an average result in one mouse (l). Two-tailed independent t-test. m, Top 10 significantly enriched canonical pathways of 3xDR versus control microglial DEGs annotated by IPA (q < 0.05). n,o, 3xDR versus control microglia FC are positively correlated with ADEM (n) and DAM (o) gene sets. Gray shading represents the 95% confidence interval, and Pearson’s correlation coefficients and P values are shown at the bottom. p,q, Representative confocal image shows that OPN and AXL are up-regulated in 3xDR microglia. n = 10 mice for each group. Two-tailed independent t-test. Data are presented as mean ± s.d. BW, body weight; Ctrl, control; MO, months old; NxDR, N-round depletion–repopulation; PLX5622, PLX5622-formulated diet.
