Morphometry analysis
Liver and skeletal muscle tissues were fixed in 10%
formaldehyde, embedded in paraffin,
and cut into 5 μm serial sections for staining with H&E and Oil Red O.
Histopathological changes were observed using a fluorescence microscope
system (TE2000, Nikon Japan) and representative images were presented.
Transcriptomic analysis
Total RNA was extracted from liver
tissue using TRIZOL reagent and purified using the NanoPhotometer®
spectrophotometer (IMPLEN, CA, USA) to eliminate ribosomal RNA, and then
stored at -80°C prior for further analysis by NEBNext® UltraTM RNA
Library Prep Kit for Illumina® (NEB, USA) following manufacturer’s
recommendations. Sample sets were constructed with six individual
samples from C57, KK-Ay and KK-Ay+Dios group (three samples per group).
RNA-sequence analysis was conducted by RNA-sequence analysis conducted
by Beijing Novogene Genomics Technology Co. Ltd. (Beijing, China). The
Gene Ontology (GO) enrichment analysis of differentially expressed genes
was implemented by the clusterProfiler R package, in which gene length
bias was corrected. GO terms with corrected P value less than 0.05 were
considered significantly enriched by differential expressed genes.
ClusterProfiler R package was used to test the statistical enrichment of
differential expression genes in KEGG pathways.