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. 2021 May 25;218(8):e20201708. doi: 10.1084/jem.20201708

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© 2021 Chen et al.

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Figure 4. — Loss of SERCA2 and SERCA3 leads to diminished Rag expression. (A) Schematic of pMGINV with locations of PCR primers (arrows) used to amplify CJ and the plasmid backbone (Amp^R). (B) Quantitative PCR analysis of pMGINV CJ formation in plasmids recovered from WT and Atp2a2^−/− abl preB cells treated with imatinib (mean ± SD; n = 3). ***, P < 0.001. (C and D) Levels of Rag1 (C) and Rag2 (D) mRNA assayed by quantitative RT-PCR in abl preB cells treated with imatinib for 1 or 2 d. WT and Atp2a2^−/− abl preB clones (−) were assayed, as were Atp2a2^−/− abl preB clones with bulk Atp2a3 inactivation (+). Rag1 and Rag2 mRNA levels were normalized to β-actin mRNA and relative to the WT value, which was set at 1. Rag1 and Rag2 mRNAs were not detected at significant levels in cells not treated with imatinib (mean ± SD; n = 3). **, P < 0.01; ***, P < 0.001 compared with WT. (E) Western blot analysis for RAG1 and actin in WT and Atp2a2^−/− abl preB cell clones (−) and cells bulk Atp2a2 (gAtp2a2) or Atp2a3 (gAtp2a3) inactivated and treated with imatinib for 1 or 2 d (n = 1 for 1 d, n = 2 for 2 d).