Fig. 3. Spliced isoforms of Apg16L. (A) Alternative splicing of Apg16L mRNA. The 20 exons are indicated by numbers above the line representing Apg16L. Alternatively spliced exons in Apg16L and Apg16Lß are indicated as broken lines. The positions of the primers (within exons 6 and 10) used in C are indicated by arrows. Corresponding domain structures are shown as in Fig. 2C. (B) Expression of Apg16L in tissues and cell lines. Tissue homogenates were prepared from mouse liver (lane 1), brain (lane 2), the gastrocnemius muscle (lane 3) and kidney (lane 4). Total cell lysates were also prepared from ES cells (lane 5) and HeLa cells transiently transfected with either vector alone (lane 6), Apg16L (lane 7) or FLAG-tagged Apg16L (lane 8). The mobility of the three isoforms is indicated. (C) Reverse-transcription-PCR analysis of Apg16L mRNA. Total RNA was isolated from mouse liver (lane 1), brain (lane 2), kidney (lane 3), ES cells (lane 4) and HeLa cells (lane 5), and reverse-transcribed into cDNA. A fragment of the Apg16L cDNA corresponding to exons 6-10 was amplified using the primers indicated in A. The cDNA sequences of mouse Apg16L, Apg16Lß and Apg16L have been deposited in the DDBJ/EMBL/GenBank databases under accession numbers AB087879, AB087880 and AB087881, respectively.