Semi-quantitative RT-PCR for Xbp1 mRNA splicing

2 μl of cDNA was added to 0.2 μM of forward and reverse primers (Hs_XBP1_Fwd: 5’ -GGAGTTAAGACAGCGCTTGG- 3’; Hs_XBP1_Rev: 5’ -ACTGGGTCCAAGTTGTCCAG-3’), 0.2 mM of each dNTP, 0.5 units of Taq DNA polymerase (Thermo Scientific). The reaction was set at an annealing temperature of 60.5°C with an extension time of 30 s for 26 cycles. The products were then visualized on a 3% agarose gel (comprised of a 1:1 mixture of low-melting point agarose and standard agarose) stained by 1:10000 SybrSAFE.