Expression of the full-length iPLA2α message in 832/12 INS-1 cells. Following total RNA isolation from 832/13 INS-1 cells, cDNA was prepared from 6 μg of RNA by RT reaction. PCR analyses were then performed for iPLA2α using five overlapping primer sets (1–5), described in the Materials and Methods. (A) Template of the iPLA2β message sequence and the regions targeted for amplification by the five primer sets. (B) PCR was performed in the presence of each primer set separately, and the resulting reaction products were analyzed on 1.0% agarose gels and visualized by ethidium bromide staining. The expected product size (bp) with each primer set is as follows: (1) 523, (2) 589, (3) 586, (4) 597, and (5) 336. The PCR bands 4a and 5a were nonspecific products whose sequences had no homology with the iPLA2β mRNA sequence.