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