Project description:MEF from p8 knock-out mice were immortalized with rasV12/E1A and infected with an empty or a p8-overexpressing retroviral vector as described. Total RNA was isolated and gene expression profile was studied.
Project description:The broilers were randomly allotted to four treatment groups (Con, DEX, P8, and DEX+P8 groups) with 10 replicates per group (10 broilers per replicate). Broilers in the Con and DEX groups were fed a basal diet. Broilers in the P8 and DEX+P8 groups were fed a basal diet containing 1 × 108 CFU/g P8. At 16 days of age, broilers in the DEX and DEX+P8 groups were injected with 3 mg/kg body weight DEX (200 μL), whereas broilers in the Con group were injected with an equal volume of saline.
Project description:Purpose - To determine transcriptomic variations in the mouse cochlea in response to three different transcription factor overexpression conditions and no induction control and to determine transcriptomic and accessible chromatin variations in the mouse cochlea between postnatal day 1 (P1) and postnatal day 8 (P8) stages of development. Methods - SC RNA seq performed on FACS enriched cells obtained from experimental animals (Sox9CreER; Rosa26A/GA/GAP/+; Rosa26 tdtomato). Same samples at two timepoints 1 week (P8) and 2 week (P15) ages. scMultiome ATAC + Gene Expression (10x Genomics) experiments on enzymatically dissociated and unenriched cochlear nuclei from wildtype mice in a mixed background of CD1 and FVB/NJ, and C57BL/6. Samples were collected at P1 and P8. Gene expression data was used to annotate clusters. Chromatin accessibility determined by ATAC was compared between P1 and P8 stages. Results - The SC RNA seq highlighted differential transcriptomic changes between the two ages and the three overexpression conditions. Hair cell loci become less epigenetically accessible in supporting cells and GER cells between P1 and P8. Conclusions - Our study highlights the variability in transcriptomic response to three transcription factor combination mediated reprogramming cues at two different ages. The SC RNA seq confirmed a variety of our biological observations with regard to the hair cell reprogramming process in the mouse cochlea. We also demonstrate variability in chromatin accessibility between P1 and P8, which may explain why GER and supporting cells of the cochlea become more resistant to transcription factor reprogramming into hair cells during the first postnatal week.
