Figure 7

SND1 regulates Ccl19 splicing. (A) RNA from the adipose tissue of control or AdipoQ-LPL mice was subjected to PCR with primers located in exon 1 and 4 of Ccl19. The PCR products were resolved on an agarose gel and then sequenced. The positions of full-length Ccl19 and the shorter isoform, which was found to lack exon 2 by DNA sequencing, are indicated. (B) The ratio of full-length Ccl19 to Ccl19 (1,3,4) was calculated. The data are represented as the mean ± SEM (n = 3; **P < 0.01; two-tailed unpaired Student’s t-test). (C) A diagram showing the four exons of Ccl19 and alternative usage of exon 2. (D) Translation of the full-length and Ccl19 (1,3,4) reveals that when exon 2 is missing, the reading frame shifts and a premature stop codon is present. The arrow indicates the position of the amino acid affected by the frame shift. (E) Inset: induction of two Ccl19 isoforms in 3T3-L1 adipocytes by TNFα. The ratio of exon 2 to exon 1 in 3T3-L1 adipocytes treated with scrambled or Snd1 siRNA and the TNFα were calculated by real time RT-PCR with exon specific primers and Ccl19 plasmid DNA to construct the standard curve. The data are represented as the mean ± SEM (n = 6; *P < 0.05; two-tailed unpaired Student’s t-test).