Fig. 2
Analysis of Dlg2 isoforms expressed in IFNβ/YFP+-producing pDCs. (a) Gene architecture and alternatively spliced transcripts of the Dlg2 gene. (a, upper graph) Genomic position of Dlg2 gene is presented. Exons are shown as vertical bars and introns as thin horizontal lines. Introns and exons are drawn to scale. Smaller exons (less than 1 point line space) are not to scale. (a, lower graph) Exons are shown as boxes and are drawn to scale. Exons are named or numbered as indicated. Alternative exon-exon-junctions are indicated with connecting lines. Grey boxes show protein coding regions whereas empty boxes represent untranslated mRNA regions. (b) RT-PCR of Dlg2 N-terminal isoforms in IFNβ/YFP+ and IFNβ/YFP— BM-derived pDCs (upper and middle panel). Flt3L cultures from six pooled IFNβmob/mob mice were stimulated with CpG for 6 h and FACS-sorted for YFP+ (pDC+) and YFP— pDCs (pDC—). Naïve brain from C57BL/6 N mice was used as positive and not reversely transcribed RNA from YFP+ pDCs as negative controls (Control). Lower panel shows Gapdh and Ifnb expression in the respective cDNA samples indicating successful stimulation and sorting of pDCs as well as equal template amounts. (c) SH3-GUK linker isoforms of Dlg2 in RT-PCR. SH3-GUK region was amplified using cDNA samples as described above in (b). (d) Restriction analysis of Dlg2 clones generated after 5´-RACE PCR. Empty vector or selected 5´-RACE clones were digested either with EcoRI (left), HindIII (middle), or with both restriction enzymes (right). Lower panel shows the plasmid maps for the clones shown in the upper panel. (e) Exon-intron structure of the Dlg2 isoforms expressed in pDCs. Exons are shown as boxes and are drawn to scale as shown in A (lower part)