Cspd reagent

Northern blot analysis of β-globin mRNA was performed as previously described (30 (link)), using digoxygenin-labeled antisense probes and alkaline phosphatase-coupled anti-digoxygenin antibody (Roche). Blots were developed with the LAS 3000 imaging system (Fujifilm) using CSPD reagent (Roche). For RT-qPCR 500 ng of total RNA was reverse-transcribed using oligo(dT)₁₈ primers (Eurofins) and RevertAid Reverse Transcriptase (Thermo Fisher Scientific). qPCRs were carried out as described (13 (link)), using TaqMan assays (Life Technologies, assay-ID Hs00230071_m1 for NFKBIZ mRNA, Hs00174131_m1 for IL6 mRNA, Hs99999905_m1 for GAPDH mRNA, Hs00262018_m1 for NFKBID mRNA, Hs00153283_m1 for NFKBIA mRNA and custom-made assays for rabbit β-globin and firefly luciferase mRNAs) or SybrGreen-based detection (Life Technologies) for mRNAs using primers as follows: GFP: sense 5′-TGCAGTGCTTCAGCCGCTAC-3′, antisense 5′-TCGCCCTCGAACTTCACCTC-3′; Renilla luciferase: sense 5′-CGTCGTGCCTCACATCGAGC-3′, antisense 5′-GCTCGAACCAAGCGGTGAGG-3′; MCPIP1: sense 5′-GGCTGGACAGAGGGAGGATT-3′, antisense 5′-TGACCCACTGAGGCAGACAG-3′; in vitro-transcribed CrPV IRES RNA: sense 5′-GCCTAATACGACTCACTATAGGGTTTAGTGAACCGTGGATC-3′, antisense 5′-GGCGGATCCTGTATCTTGAAATGTAGCAGGTAAA-3′; in vitro-transcribed TSE RNA: sense 5′-TTGGAGCCTGGCTAGCAACA-3′, antisense 5′-TTGCCAATATAAGGCAAATGGTCT-3′.