Journal: Cell communication and signaling : CCS

Article Title: Cold-blooded vertebrates evolved regulatory B cells to participate in inflammatory diseases.

doi: 10.1186/s12964-025-02311-y

Figure Lengend Snippet: Fig. 1 Teleost CD25L+B cells have the ability to produce IL-10 and IL-12p35. A Cell sorting and double check of cell purity of CD25L− B and CD25L+ B cells from grass carp PBLs. B Giemsa’s staining and TEM observation of the morphology and ultrastructure of CD25L− B and CD25L+ B cells (N, nucleus; yellow arrow, organelle). Scale bar, 10 μm (left); 1 μm (right). C Analysis of cell size and granularity of two B cell subsets. n = 12 each. D Significantly en riched gene sets related to cytokine production in CD25L+ B cells (NES, normalized enrichment score; FDR, false discovery rate). E Heatmap analysis of phenotypic DEGs related to cytokine production in the two B cell populations. n = 3 each. F Scatterplots of DEGs related to the key transcription factors and cytokines in CD25L+ B versus CD25L− B (data adjusted p < 0.01,|log2FC| > 1; red, upregulated genes; blue, downregulated genes). n = 3 each. G, H Relative mRNA (G) and protein (H) levels of the indicated molecules in CD25L− B cells and CD25L+ B cells. Total leucocytes served as controls. n = 3 each. I Immunofluorescence co-localization of IL-10 (red) or IL-12p35 (red) combined with B cells stained with IgM (green) and CD25L (magenta). Scale bar, 5 μm. Data acquisition required three independent tests (mean ± SEM). n represents biological replicates. Unpaired t-test was used for (C) and one-way ANOVA was used for (G) and (H). ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. PBLs, peripheral blood leukocytes; DEGs, differentially expressed genes; TEM, transmission electron microscope

Article Snippet: After incubation, cells were stained with 2 μg/ml Cy3 goat anti rabbit IgG and 2 μg/ml FITC goat anti mouse IgG (#405305, BioLegend) for 2 h at room temperature, then stained with DAPI and imaged by inverted confocal microscope (Nikon N-STORM) with deconvolution.

Techniques: FACS, Staining, Immunofluorescence, Transmission Assay, Microscopy

Journal: Scientific reports

Article Title: Histamine stimulates human microglia to alter cellular prion protein expression via the HRH2 histamine receptor.

doi: 10.1038/s41598-024-75982-1

Figure Lengend Snippet: Fig. 3. Histamine stimulates HMC3 cells to alter cytokine production and intracellular calcium levels, but not metabolic activity or Iba-1 expression. (A-B) HMC3 cells were stimulated with (A) histamine (0.1 µM – 1000 µM) or (B) LPS or left untreated for 24 hours and metabolic activity was measured by a reduction in XTT. The data are presented as the means ± SEMs (N = 6). (C-D) HMC3 cells were stimulated by histamine (10 µM, 100 µM, or 1000 µM), LPS (1 µg/mL), or left untreated for 24 hours, and (C) IL-8 and (D) IL-6 were measured by sandwich ELISA. Data are presented as the mean ± SEM (N = 9) and statistical significance was measured via one-way ANOVA and Dunnett’s multiple comparison post-hoc analysis relative to untreated (UT) cells. p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***), p ≤ 0.0001 (****). (N = 4). (E) HMC3 cells were stimulated by histamine (100 µM) (blue) or 4-bromo-A23187 (1 µM) (orange) and intracellular calcium was measured via fura-2 AM dye which increases the 340/380 fluorescence ratio when bound to calcium ions. (F-H) Fluorescence microscopy images of HMC3 cells labelled with Hoechst 33342 (Hoechst) (blue) and Iba-1 (red) under (F) untreated conditions and following 24 hours treatment with (G) LPS and (H) histamine. Images are representative of four independent experiments using a 20× objective and a Leica Stellaris 5 microscope.

Article Snippet: Images were acquired using an Olympus IX81 inverted microscope with a 20× objective and a Leica Stellaris 5 laser scanning confocal microscope with a 60× oil objective.

Techniques: Activity Assay, Expressing, Sandwich ELISA, Comparison, Fluorescence, Microscopy

Journal: Scientific reports

Article Title: Histamine stimulates human microglia to alter cellular prion protein expression via the HRH2 histamine receptor.

doi: 10.1038/s41598-024-75982-1

Figure Lengend Snippet: Fig. 7. Immunofluorescence detection of surface PrPCexpression in HMC3 cells treated with histamine and LPS. Representative fluorescence microscopy images of HMC3 labelled with Hoechst 33342 (blue) and anti- PrPC POM2 (red) under (A) untreated conditions or after 24 hours of stimulation by (B) histamine or (C) LPS. Images are representative of three independent experiments using a 20× objective and a Leica Stellaris 5 microscope.

Article Snippet: Images were acquired using an Olympus IX81 inverted microscope with a 20× objective and a Leica Stellaris 5 laser scanning confocal microscope with a 60× oil objective.

Techniques: Immunofluorescence, Fluorescence, Microscopy