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. Author manuscript; available in PMC: 2022 Dec 14.
Published in final edited form as: Nat Cardiovasc Res. 2022 Nov 11;1:1056–1071. doi: 10.1038/s44161-022-00155-0

Figure 4. Impact of global GPR55 deficiency on B cell transcriptome and mitochondrial content.

Figure 4

(A) Volcano plot of differentially expressed genes (DEGs) in Apoe-/-Gpr55-/- versus Apoe-/- splenic CD19+ cells after 4 weeks WD. The x-axis shows log2 fold changes between genotypes. The y-axis plots negative log10 P-values computed by DEseq2 (based on a generalized linear model) and corrected for multiple comparisons according to the Benjamini-Hochberg FDR; relevant statistically upregulated (blue) and downregulated (red) genes are highlighted; n=6). (B) Confirmation of the main DEGs in splenic B cells of Apoe-/-Gpr55-/- compared to Apoe-/- mice after 4 weeks WD by qPCR (n=5; p=0.05). (C) Chord diagram showing the main regulated genes and pathways in splenic Apoe-/-Gpr55-/- versus Apoe-/- B cells, splenic PCs and circulatory plasmablasts, based on RNA-sequencing. (D) Splenic B cell Gpr55 mRNA expression (qPCR) correlated with Fcer2a (CD23 encoding gene) mRNA expression and (E) splenic B cell Gpr55 mRNA expression (qPCR) correlated with Ptcarpc (LPAP encoding gene) mRNA expression (n=20 Apoe-/- mice). (F) Mean fluorescence intensity (MFI) of CD23 on splenic Apoe-/- B cells measured by flow cytometry after LPI treatment (1 μM; n=4). (G) Flow cytometric analysis of CD23 MFI on splenic B cell subsets of Apoe-/- and Apoe-/-Gpr55-/- mice after 4 weeks WD (n=8-10; ***p<0.0001). (H) Mitochondrial content of splenic GC B cells (n=6-7; ***p < 0.0001). (I) Mitochondrial content of splenic PCs (n=6-7; ***p < 0.0001). (J) Mitochondrial content of bone marrow long-lived PCs (n=6-7; ***p < 0.001). Data shown in D-J were combined from 2 independent experiments, and each dot represent one biologically independent mouse sample for all figure panels (mean ± SEM). Two-sided unpaired Student’s t-test or one-way-ANOVA followed by a post-hoc Newman–Keuls multiple-comparison test was used to determine the significant differences. Bivariate correlations were analyzed by Spearman’s rank correlation test. FO, follicular; GC, germinal center; LPI, lysophosphatidylinositol; MFI, mean fluorescence intensity; MZ, marginal zone; PC, plasma cell.