Nfil3, a target of the NACA transcriptional coregulator, affects osteoblast and osteocyte gene expression differentially.

Purpose: Identify PTH-responsive genes in MC3T3-E1 cells. The aim of the study is to identify novel gene targets that are reugulated by PTH signaling in bone, elicting its anabolic effect in osteoblasts. Methods: mRNA expression profiles were compared between Vehicle and PTH(1-34)-treated MC3T3-1 cells, in triplicates, using Illumina HiSeq 2000 PE100 sequncing. qPCR using Taqman probes were used to validate genes of interest. Results: Sequence reads per sample were mapped to the mouse genome (mm10) and differential expression of each gene was determined. Genes that showed differential expression between Vehicle vs PTH(1-34)-treated samples and has a relevant function in bone were further characterized by knockdown strategies in MC3T3-E1 cells. Conclusion: Our study have identified a list of novel genes regulated by PTH siganling. The characterization of their transcription and function in osteoblasts and osteocytes added to our knowledge of PTH signaling in bone and other skeletal tissues. Overall design: Analysis of total RNA isolated from MC3T3-E1 treated with Vehicle or PTH(1-34).