Fig 1: Autoregulation of XBP1. hSAECs were RSV infected (MOI = 1.0) for 24 h in the presence of DMSO, KIRA8 (KIRA), or ceapin-A7 (A7) at 10 µM. A and B: Q-RT-PCR of total XBP1 (XBP1-Total) and unspliced XBP1 (XBP1u). Shown is fold change of mRNA relative to mock infection (DMSO) in n = 3 independent experiments. Recruitment of XBP1 (C) and RNA Pol II (D) to XBP1 promoter. XChIP was conducted using XBP1 or RNA Pol II-specific antibodies (XBP1, Cat. No. ab37152 at Abcam; RNA Pol II, Cat. No. ab26721 at Abcam). The XChIP-enriched genomic DNA was analyzed by Q-gPCR using XBP1 promoter-specific primers (Table 2). Data are calculated as fold change relative to uninfected hSAECs (DMSO) and plotted as means ± ranges plus all data points of duplicate independent immunoprecipitates. Note the RNA Pol II dependence on IRE1a-XBP1 signaling. **P < 0.01, ANOVA. hSAECs, human small airway epithelial cells; MOI, multiplicity of infection; Q-RT-PCR, quantitative reverse transcription polymerase chain reaction; RSV, respiratory syncytial virus; XBP1, X-box binding protein 1.
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