Journal: Cell reports

Article Title: The microglial response to inhibition of Colony-stimulating-factor-1 receptor by PLX3397 differs by sex in adult mice.

doi: 10.1016/j.celrep.2024.115176

Figure Lengend Snippet: Figure 1. Sex-dependent differences in microglia density and morphology under CSF1R inhibition (A) Representative 103 epifluorescence images (cropped from a stitched image representing the whole brain section) showing microglia immunolabeled with Iba1 in hippocampal CA1, white matter (WM), and primary visual cortex V1 in control and PLX-treated male and female mice. (B) Quantification of depletion efficacies across different brain regions as depletion/control microglia density ratios. Briefly, all control animals of each sex were averaged, and the depletion/control ratios were calculated as each PLX animal divided by the control animal average of the same sex. (C) Microglial soma size was unaltered after PLX treatment. (D) Representative 203 confocal image of microglia in V1. Outline shows how individual microglia were cropped out of the image for analysis. (E) Representative image of a thresholded microglia with overlaid concentric circles for Sholl analysis. (F–I)QuantificationofmicroglialramificationastotalShollintersectionsdependingontreatmentandsexinV1(F);CA1,includingSO,SP,andSR(G);SLM(H);andWM(I). n = 6–7 mice/sex/treatment (B and C), n = 3–5 mice/sex/treatment (F–I); two-way ANOVA with Bonferroni correction, *p < 0.05, **p < 0.01, ***p < 0.001. Scale bars: 200 mm. All data are expressed as the mean ± SEM. See also Figures S1–S3.

Article Snippet: After PLX treatment, CSF1R expression decreased in both Galectin-3+ and Galectin-3 microglia in both sexes (Figures 5H and 5I), and Galectin-3+ microglia had higher CSF1R expression than Galectin-3 microglia in males (Figure 5I), while expression normalized between the two populations in females (Figure 5H).

Techniques: Inhibition, Immunolabeling, Control

Journal: Cell reports

Article Title: The microglial response to inhibition of Colony-stimulating-factor-1 receptor by PLX3397 differs by sex in adult mice.

doi: 10.1016/j.celrep.2024.115176

Figure Lengend Snippet: Figure 3. Sex-dependent differences in microglial transcriptomic landscape during CSF1R inhibition (A) Volcano plot showing DEGs between female and male microglia in control animals. (B) Enrichment analysis revealed upregulation of immune-related pathways in control male microglia compared to female. The small number of upregulated DEGs in females compared to males precluded an enrichment analysis. (C) Volcano plot showing DEGs between female and male microglia after 7 days of PLX treatment. (D) After PLX treatment, upregulated DEGs in females were enriched in pathways related to autophagy and protein homeostasis. (E) After PLX treatment, upregulated DEGs in males were enriched in mitochondrial oxidative phosphorylation and mitotic processes. n = 4 mice/sex/treatment; adjusted p cutoff <0.05. See also Figures S4–S6 and Tables S2 and S5.

Article Snippet: After PLX treatment, CSF1R expression decreased in both Galectin-3+ and Galectin-3 microglia in both sexes (Figures 5H and 5I), and Galectin-3+ microglia had higher CSF1R expression than Galectin-3 microglia in males (Figure 5I), while expression normalized between the two populations in females (Figure 5H).

Techniques: Inhibition, Control, Phospho-proteomics

Journal: Cell reports

Article Title: The microglial response to inhibition of Colony-stimulating-factor-1 receptor by PLX3397 differs by sex in adult mice.

doi: 10.1016/j.celrep.2024.115176

Figure Lengend Snippet: Figure 4. Sex-dependent differences in microglial Galectin-3 expression at baseline and under CSF1R inhibition (A and B) Representative 203 confocal images showing microglia immunolabeled with Iba1 (green) and Galectin-3 (magenta) in control (A) and PLX-treated mice (B). For ease of visualization, representative images were subjected to altered brightness/contrast that masks the different Galectin-3 fluorescence intensity. (C) The percentage of Galectin-3+ microglia was higher in females compared to males in both cortex (left) and hippocampus (right), although the effect did not reach statistical significance. (D and E) PLX treatment increased the proportion (D) and density (E) of Galectin-3+ microglia in both males and females, also increasing the level of expression in individual microglia. n = 6–8 mice/sex/treatment (C), n = 3–8 mice/sex/dose (D and E), Student’s t test (C), two-way ANOVA with Bonferroni correction (D and E), p values are reported in Tables S4 and S5. Scale bar: 200 mm. All data are expressed as the mean ± SEM. See also Figure S6.

Article Snippet: After PLX treatment, CSF1R expression decreased in both Galectin-3+ and Galectin-3 microglia in both sexes (Figures 5H and 5I), and Galectin-3+ microglia had higher CSF1R expression than Galectin-3 microglia in males (Figure 5I), while expression normalized between the two populations in females (Figure 5H).

Techniques: Expressing, Inhibition, Immunolabeling, Control

Journal: Cell reports

Article Title: The microglial response to inhibition of Colony-stimulating-factor-1 receptor by PLX3397 differs by sex in adult mice.

doi: 10.1016/j.celrep.2024.115176

Figure Lengend Snippet: Figure 5. Expression patterns of Galectin-3+ microglia at baseline and under CSF1R inhibition (A) Experimental paradigm depicting duration of PLX treatment and subsequent microglia gating and analysis. (B) PLX treatment increases the proportion of microglia expressing Galectin-3 to a greater extent in males compared to females. (C) PLX treatment increases the expression levels of Galectin-3 in Galectin-3+ microglia to a greater extent in males compared to females. (D–F) PLX treatment led to increased mitochondrial mass in female and male microglia, as estimated by expression of MitoTracker green. This effect was larger in males than in females (D). In both control and PLX groups, Galectin-3+ microglia appear to have a higher mitochondrial mass compared to Galectin-3 microglia in females (E) and males (F).

Article Snippet: After PLX treatment, CSF1R expression decreased in both Galectin-3+ and Galectin-3 microglia in both sexes (Figures 5H and 5I), and Galectin-3+ microglia had higher CSF1R expression than Galectin-3 microglia in males (Figure 5I), while expression normalized between the two populations in females (Figure 5H).

Techniques: Expressing, Inhibition, Control

Journal: Frontiers in immunology

Article Title: Interleukins 4 and 21 Protect Anti-IgM Induced Cell Death in Ramos B Cells: Implication for Autoimmune Diseases.

doi: 10.3389/fimmu.2022.919854

Figure Lengend Snippet: FIGURE 9 | IL-21 rescued CD40L and anti-IgM induced cell death in human primary B cells. (A) Representative graphs of CFSE labelling cells showed no differentiation was induced by any stimulatory factors in 3 days. (B) Representative graphs showed that IL-21 could suppress cell death induced by CD40L and anti- IgM in terms of increased number of DAPI- cells. (C) Representative graphs showed that IL-21 was able to induce CD38+ B cell differentiation both in the absence and presence of CD40L.

Article Snippet: Cells were then seeded into 96-well round bottom plate at the density of 2E5 cells/100μl, and then challenged with 50 ng/ml IL-21 and 1 μg/ml CD40L (#6420-CL/CF, R&D Systems, Minneapolis, USA) or 2 μg/ml anti-IgM.

Techniques: Cell Differentiation