Figure 6.
Figure 6. A category of pDCs residing in bone marrow expresses low levels of RAG1, as well as B-lineage–associated genes, and undergoes DH-JH rearrangements. (A) An uncompensated 2-parameter procedure was used to discriminate and sort GFP+ and GFP- subsets of CD45R/B220+CD19--enriched cells from BM of RAG1/GFP knock-in mice. The purified populations were then stained with Ly-6C and CD11c. The boxes in the bottom 2 histograms indicate gates used for an additional sorting of B220+CD19-Ly-6C+CD11cLoGFP+ and B220+CD19-Ly-6C+CD11cLoGFP- pDC subsets. (B) RT-PCR analyses of the indicated genes were performed with cDNA prepared from these 2 sorted subsets and compared with B220+CD19+ CD43+CD24+ pro-B/large pre-B cells or to B220-CD11c+ DCs, while β-actin was used as a loading control. (C) Genomic DNA from these same fractions was evaluated for Ig DHJH and VH-DJH rearrangements and germ-line loci by PCR. Specific products with the expected sizes are labeled with respect to rearrangement to J1, J2, or J3. Alpha actin was used as a control for genome representation.

A category of pDCs residing in bone marrow expresses low levels of RAG1, as well as B-lineage–associated genes, and undergoes DH-JH rearrangements. (A) An uncompensated 2-parameter procedure was used to discriminate and sort GFP+ and GFP- subsets of CD45R/B220+CD19--enriched cells from BM of RAG1/GFP knock-in mice. The purified populations were then stained with Ly-6C and CD11c. The boxes in the bottom 2 histograms indicate gates used for an additional sorting of B220+CD19-Ly-6C+CD11cLoGFP+ and B220+CD19-Ly-6C+CD11cLoGFP- pDC subsets. (B) RT-PCR analyses of the indicated genes were performed with cDNA prepared from these 2 sorted subsets and compared with B220+CD19+ CD43+CD24+ pro-B/large pre-B cells or to B220-CD11c+ DCs, while β-actin was used as a loading control. (C) Genomic DNA from these same fractions was evaluated for Ig DHJH and VH-DJH rearrangements and germ-line loci by PCR. Specific products with the expected sizes are labeled with respect to rearrangement to J1, J2, or J3. Alpha actin was used as a control for genome representation.

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