Fig 1: Macrophages deficient in Kdm2a increase energy expenditure and adaptive thermogenesis.A Results for real-time recording of RER (left) and quantitative data (right) collected from four mice in each group for 24 h following 16 weeks of HFD induction (n = 4 per group). B Food intake (g) in WT and KO mice following 16 weeks of HFD induction measured in metabolic cages (n = 4 per group). RT-PCR results for analysis of thermogenic genes Cox5a, Ucp1, Cox7a, and Cox8b in BAT (C) and scWAT (D) from HFD challenged WT and KO mice (n = 6 for each group). E RT-PCR results for analysis of Ucp1 in the epWAT following 16 weeks of HFD challenge (n = 6 for each group). F Daily rectal temperature of mice housed under 4 °C condition (n = 6 per group). G Analysis of the weight for BAT collected from WT and KO mice after cold exposure (n = 6 per group). H Representative western blot results (left) and bar graphs (right) showing the expression levels for BAT TH and Ucp1 (n = 4 per group). I Relative mRNA abundance of Ucp1 in BAT of mice with (n = 6/group) or without cold stress (n = 3/group). Representative images of H&E staining (J) and Ucp1 IHC (K) of BAT sections with or without cold exposure (six images per mouse). Representative images of H&E staining (L) and Ucp1 IHC (M) of scWAT sections (six images per mouse). Scale bars: 50 µm (J–M). Original magnification: ×400 (J–M). Values are expressed as mean ± SEM, and unpaired Student’s t test was used for data analysis. *P < 0.05; **P < 0.01; ***P < 0.001. ns not significant.
Fig 2: The mRNA expression levels of positive and negative genes related to regulate oogenesis and folliculogenesis in GV-stage oocytes. Four-week-old ovaries from Kdm2a cKO (cKO) and control (Con) mice were used to collect GV-stage oocytes; then, RT-qPCR was used to determine the levels of transcripts positively (A) and negatively (B) associated with oocyte and follicle development. n = 5 females/group. * Significant difference (p < 0.05).
Fig 3: Subcellular localization and expression of Kdm2a during oocyte meiotic maturation. (A,B) Immunofluorescence analysis was utilized to track the spatial profile of KDM2a in GV, GVBD, and MII stages oocytes. Oocyte were labeled with anti-KDM2a followed by appropriate Alexa Fluor-conjugated secondary antibodies (green) and were counterstained with the nuclear stain PI (red). (C,D) Protein levels of KDM2a in oocytes at GV, GVBD, and MII stages were examined by western blot. (E) RT-qPCR analysis of Kdm2a mRNA relative expression levels in oocytes. The different superscript letter showed significant difference (p < 0.05). Scale bars, 50 µm.
Fig 4: KDM2A promotes Treg cell proliferation and glioma immune tolerance by upregulating JAG1 through affecting H3K4me3 in the JAG1 promoter.A JAG1 expression in glioma tissues (N = 40) and normal brain tissues (N = 10) determined by RT-qPCR. B JAG1 expression in glioma tissues (N = 40) and normal brain tissues (N = 10) detected by IHC. C Correlation analysis between KDM2A and JAG1 in glioma tissues (N = 40) using Pearson’s correlation coefficient. D KDM2A mRNA and protein expression in LN229 and A172 cells treated with si-KDM2A-1 and si-KDM2A-2 determined by RT-qPCR and Western blot analysis. E The enrichment of KDM2A and H3K4me3 on the JAG1 promoter determined by ChIP. LN229 and A172 cells were treated with si-NC + oe-NC, si-KDM2A-1 + oe-NC, or si-KDM2A-1 + oe-JAG1. F JAG1 and KDM2A mRNA and protein expression in LN229 and A172 cells determined by RT-qPCR and Western blot analysis. G LN229 and A172 cell proliferation detected by CCK-8 assay. H Ratio of CD4+CD25+Foxp3+ cells in CD4+ cells co-cultured with LN229 and A172 cells analyzed by flow cytometry. *p < 0.05. Data were shown as the mean ± standard deviation from. Statistical comparisons were performed using unpaired t-test when only two groups were compared or by one-way ANOVA with Tukey’s post hoc test when more than two groups were compared. Variables were analyzed at different time points using two-way ANOVA with Bonferroni post hoc test. Cell experiments were repeated three times.
Fig 5: Analysis of H3K36me1/2/3 levels in the GV-, GVBD-, and MII-stage oocytes of control and Kdm2a cKO mice. (A) Analysis of H3K36me1/2/3 levels by IF. Oocytes were labeled with anti-H3K36me1/2/3 followed by appropriate Alexa Fluor-conjugated secondary antibodies (green) and were counterstained with the nuclear stain PI (red). (B–D) The H3K36me1/2/3 fluorescence intensity in oocytes of control and Kdm2a cKO mice by image pro-plus 6.0. * Significant difference (p < 0.05) and ** Extremely significant difference (p < 0.01) from the Kdm2a cKO group compared with the control group.
Supplier Page from Abcam for Anti-KDM2A antibody [EPR18602]