Journal: Signal Transduction and Targeted Therapy
Article Title: Ring finger protein 213 regulates B-cell receptor signaling, metabolism, and development in B lymphocytes
doi: 10.1038/s41392-026-02575-x
Figure Lengend Snippet: RNF213 regulated antigen presentation, NFκB and STAT signaling activation, and metabolism in splenic B lymphocytes. a GSEA identified the significantly downregulated KEGG pathways in KO B cells. b GSEA revealed enrichment of the fatty acid metabolism hallmark pathway in B cells. c Western blotting of pP85, pP55, pAKT, pmTOR, pS6, pFOXO1 and their respective total proteins in B cells stimulated with or without anti-BCR for 5 min. Statistical analyses of phosphorylated proteins are shown. d , e Flow cytometry and statistical analysis of ROS ( d ) and mitochondrial membrane potential (e ) levels in WT (n = 3) and KO (n = 3) B cells stimulated with anti-BCR for 3 h or in WT (n = 5) and KO (n = 4) B cells without stimulation. f , g Energy metabolism in anti-BCR-stimulated B cells from WT and KO mice was examined via Seahorse technology. Glycolysis and mitochondrial OXPHOS were evaluated by the ECAR (n = 5) (f ) and OCR (n = 4) ( g ), respectively. Left, representative dynamic curves. Right, statistical analysis of each metabolic phase. Each symbol represents the mean ± SEM of cells (left) or one well of cells, with bars denoting the means ± SEMs (right). Data are from one representative of three independent experiments in ( c–g ). Each symbol represents one mouse, with bars denoting the means ± SEMs in ( c-e ). Statistical analyses were performed via unpaired two-tailed Student’s t tests. The following symbols indicate statistical significance: * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant
Article Snippet: The cell lysates were examined via SDS-PAGE, and immunoblotting was performed with the following antibodies: anti-pSYK (Cell Signaling Technology, CST, 2710S), anti-SYK (CST, 13198S), anti-pCD19 (CST, 3571S), anti-CD19 (CST, 90176S), anti-β-actin (Proteintech, 66009-1-Ig), anti-pNFκB pP65 (CST, 3033S), anti-NFκB P65 (CST, 4764S), anti-pSTAT1 (CST, 9167S), anti-STAT1 (CST, 14994S), anti-pSTAT3 (CST, 9145S), anti-STAT3 (CST, 9139S), anti-pSTAT5 (CST, 4322S), anti-STAT5 (CST, 94205S), anti-pPI3K pP85/pP55 (CST, 4228 L), anti-PI3K P85 (CST, 4292S), anti-pAKT (CST, 4060 L), anti-AKT (CST, 9272S), anti-pmTOR (CST, 5536S), anti-mTOR (CST, 2983S), anti-pS6 (CST, 4858 L), anti-S6 (CST, 2217S), anti-pFOXO1 (CST, 9461S), anti-FOXO1 (CST, 2880S), anti-CTCF (ABclonal, A19588), anti-SPIB (CST, 14337S), anti-ETS1 (ZenBio, R382074), anti-GABPA (HuaBio, HA500187 ), anti-FLI1 (HuaBio, HA500149 ), anti-PU.1 (ABclonal, A20461), anti-GAPDH (Proteintech, 60004-1-Ig), anti-PIK3C3 (ABclonal, A12295), anti-PTEN (CST, 9188S), anti-pSHIP-1 (CST, 3941S), SHIP-1 (ABclonal, A3571), anti-NOTCH2 (ABclonal, A24877), anti-pHSL (CST, 4126S), anti-HSL (CST, 4107S), anti-ATGL (CST, 2138S), anti-LC3 I/II (ABclonal, A5618), IκBα (ABclonal, A19714), pIκBα (SelleckChem, F2237), pIKKα/β (CST, 2697S), pP38 (SelleckChem, F0159), HSP90 (ABclonal, A5027), pJNK1/2 (ABclonal, AP0473), pERK1/2 (ABclonal, AP0472).
Techniques: Immunopeptidomics, Activation Assay, Western Blot, Flow Cytometry, Membrane, Two Tailed Test
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