Transcriptomic profiling of zebrafish livers with nAtf6 overexpression at collected at 78 and 120 hpf

Activting transcription factor 6 (Atf6) is one of three main mediators of the Unfolded Protein Response and it gets cleaved cleaved in the Golgi and its N-terminal consisting of the bZip domain translocates into the nucleus to generate a transcriptional response (nAtf6). This study carries out RNAseq on an established transgenic overexpression of nAtf6 in zebrafish hepatocytes ( tg(fabp10:nAtf6-cherry; cmlc2:GFP)-C allele). Samples were collected at 78 and 120 hpf to detect gene expression changes at early and late time points after nAtf6 is overexpressed. Overall design: We used two transgenic lines: Tg(fabp10a: CAAX-EGFP) to use as a marker of the liver for dissction and tg(fabp10:nAtf6-cherry; cmlc2:GFP)-C allele to overexpress nAtf6 specifically in hepatocytes.This experiment was conducted by natural spawnings between tg:(fabp10:nAtf6-cherry; cmlc2:GFP)-C and tg(fabp10a:CAAX-EGFP ). Using GFP to guide microdissscion of 20-25 livers at 78 hpf and 120 hpf . Total RNA isolated using TRIZOL and treated by DNAse I. Libraries were prepared using RiboZero (for 120 hpf samples) or polyA selection (for 78 hpf samples). Samples from 2 clutches collected at 78 hpf and 3 clutches at 120 hpf. Samples were sequenced on NextSeq550 (Illumina)