INS 832/13 cells were harvested after transfection with siRNAs or treatment with tunicamycin (final concentration of 2.5 μg/ml), total RNA was isolated using PureLink RNA Mini Kits (Invitrogen no. 12183018A), and cDNA was synthesized using First Strand cDNA Synthesis Kits (OriGene no. NP100042). The following primers were used to amplify spliced and unspliced xbp-1 transcripts: 5′-ATTCTGACGCTGTTGCCTCT-3′ (forward) and 5′-CTCTGGGGAAGGACATTTGA-3′ (reverse).

PCR products were subjected to 2.5% agarose gel electrophoresis. The size difference between products amplified from spliced and unspliced xbp-1 cDNA templates is 26 nucleotides.

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