Fig 1: Increased CRAC channel in B cells from patients with LN. PBMCs were isolated from patients with lupus nephritis (LN), rheumatoid arthritis (RA), primary Sjogren’s syndrome (pSS), or healthy controls (HC). Orai1, Orai2, and STIM2 expression in B cells was measured by flow cytometry or Western blot. (A–F) ORAI1, ORAI2, and STIM2 expression in B cells from LN, RA, pSS, or HC was measured by flow cytometry. Representative histograms were gated on CD19+ B cells. Mean fluorescence intensity (MFI) of ORAI1, ORAI2, and STIM2 from LN (n = 20), RA (n = 10), pSS (n = 10), or HC (n = 10) were summarized. (G) CD19+ B cells were isolated from LN or HC. ORAI1, ORAI2, and STIM2 expression in B cells were measured by Western blot. GAPDH was used as internal control, and relative expression values are indicated above each lane. Representative bands are shown. (H–J) Mean fluorescence intensity (MFI) of Orai1, Orai2, and STIM2 in naive B cells (CD19+IgD+CD27−), switched memory B cells (CD19+IgD−CD27+, SM), unswitched memory B cells (CD19+IgD+CD27+, UNSM), and double-negative B cells (CD19+IgD−CD27−, DN) were analyzed by flow cytometry and are summarized in dot plot with bar plot. All data are mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 by one-way ANOVA. ns, not significant.
Fig 2: Enriched calcium signaling in B cells from patients with LN. (A–E) PBMCs were isolated from patients with lupus nephritis (LN), rheumatoid arthritis (RA), primary Sjogren’s syndrome (pSS), or healthy controls (HC). Naive B cells (CD19+IgD+CD27−), switched memory B cells (CD19+IgD−CD27+, SM), unswitched memory B cells (CD19+IgD+CD27+, UN-SM), and double-negative B cells (CD19+IgD−CD27−, DN) were analyzed by flow cytometry. (A) Gating strategy. Percentages of (B) naive B cells, (C) SM B cells, (D) UNSM B cells, and (E) DN B cells are summarized in dot plot with bar plot. LN = 40, RA = 20, pSS = 10, HC = 20. (F–J) RNA-seq data of B cell purified from peripheral blood of patients with systemic lupus erythematosus (SLE) or HC were acquired from the database (GSE118254) for gene expression and bioinformatics analysis. (F) Gene expression clustered heatmap represents the scaled average expression of top 200 DEGs. (G) Bubble plot of GO and KEGG analysis. The number of DEGs enriched in different terms are indicated by size of dots. (H) Clustered heatmap of the scaled average expression 79 DEGs in three calcium-related GO classes. (I, J) Violin plot of ORAI1, ORAI2, ORAI3, STIM1, and STIM2 expression levels. The upper, median, and lower horizontal lines of white box represent 75th, median, and 25th percentage of the gene expression distribution, respectively. All data are mean ± SEM. *p < 0.05 and **p < 0.01 by one-way ANOVA in Panel (B–E) and Student’s t-test in Panels (I, J) ns, not significant.
Supplier Page from Abcam for Anti-Orai2 antibody [EPR10043(2)] - C-terminal