Journal: Cell reports
Article Title: Transcriptional programming mediated by the histone demethylase KDM5C regulates dendritic cell population heterogeneity and function
doi: 10.1016/j.celrep.2024.114506
Figure Lengend Snippet: (A) Geometric mean fluorescence intensity (gMFI) of activation markers of Ly6C + and Ly6C − pDCs from control or Kdm5c ΔItgax mice. (B) Bone marrow chimeras as in , and gMFI of MHC II and CD40 expressed by splenic cDC subsets derived from CD45.1 + control and CD45.2 + Kdm5c ΔItgax cells 7 weeks post injection. (C) Histograms of SiglecH, Ly6C, CX3CR1, and CCR9 expression by splenic Ly6C + and Ly6C − pDCs from control and Kdm5c ΔItgax mice. (D) Heatmap of significantly different genes (false discovery rate of <0.05) between Kdm5c ΔItgax Ly6C − and Ly6C + pDCs with or without LCMV infection (20 h) of indicated pathways. (E) Heatmap of DEGs from Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways on antigen processing and presentation of control and Kdm5c Δitgax Ly6C + pDCs from mice with or without LCMV infection. (F) Ly6C + and Ly6C − pDCs were sorted from control and Kdm5c ΔItgax spleens, incubated with or without 100 μg OVA, and cultured with naive OTI CD8 or OT-II CD4 T cells for 3 days. Cell counts of OT-II CD4 co-cultured with Ly6C + pDCs (right). (G and H) Cell counts of OTI CD8 T cells and representative plots of violet proliferation dye (VPD) by CD44 of OTI CD8 T cells after co-culture with (G) Ly6C + pDCs and (H) Ly6C − pDCs. (I) Ly6C + pDC were sorted from control and Kdm5c ΔItgax mice 1 dpi with LCMV-ARM and co-cultured with P14 CD8 T cells for 3 days. Cell counts for total divided P14 CD8 T cells (top right) and for each division (bottom left). Representative plots of CD44 by VPD with gates on divided P14 CD8 T cells. (J) IFN-α production by sorted pDC subsets from control and Kdm5c ΔItgax mice stimulated with TLR9 ligand CpG ODN 1585 for 18 h (ELISA). (K) qPCR of Kdm5c from sorted splenic Ly6C + pDCs at steady state and 8 dpi with LCMV-CL13. Data are of (A, B) one experiment representative of two to five experiments (mean and SEM of 3–8 mice per experiment), (F–H) one experiment representative of two experiments (mean and SEM of 4–6 replicates per group), (I) one experiment representative of two experiments (mean and SEM of 3 per group), (J) sorted DCs pooled from three control and Kdm5c ΔItgax mice mean and SD and (K) pooled RNA from sorted pDC from six mice run in triplicate (mean and SEM). Statistical significance was determined by (H and K) unpaired t test, (A and I) multiple unpaired t test, or (B and J) one-way ANOVA. See also .
Article Snippet: Anti-mouse CX3CR1 PE (clone SA011F11) , Biolegend , 149006; RRID:AB_2564314.
Techniques: Fluorescence, Activation Assay, Control, Derivative Assay, Injection, Expressing, Infection, Incubation, Cell Culture, Co-Culture Assay, Enzyme-linked Immunosorbent Assay
