Project description:The expression of Glis3 in C3H10T1/2 cells promotes osteoblastic differentiation as indicated by the the induction of increase in alkaline phosphatase activity, an early marker of osteoblast differentiation, and increased expression of osteopontin, a late marker of osteogenesis. Glis3 acts synergistically with bone morphogenic protein 2 (BMP-2). In contrast, expression of Glis3 inhibits the induction of adipocyte differentiation. Microarray analysis identified the fibroblast growth factor 18 (FGF18) as one of the genes induced by Glis3 in C3H10T1/2 cells directly. Keywords: Glis3, osteoblast differentiation, adipocyte differentiation, FGF18, BMP2
Project description:The expression of Glis3 in C3H10T1/2 cells promotes osteoblastic differentiation as indicated by the the induction of increase in alkaline phosphatase activity, an early marker of osteoblast differentiation, and increased expression of osteopontin, a late marker of osteogenesis. Glis3 acts synergistically with bone morphogenic protein 2 (BMP-2). In contrast, expression of Glis3 inhibits the induction of adipocyte differentiation. Microarray analysis identified the fibroblast growth factor 18 (FGF18) as one of the genes induced by Glis3 in C3H10T1/2 cells directly. Experiment Overall Design: Total RNA was extracted from C3H10T1/2 cells infected with empty vector or Glis3 using Qiagen Rneasy Mini kit. Each RNA sample was amplified and converted to fluorescently labeled cRNA, either with cyanine 3 (Cy3) or cyanine 5 (Cy5), using the Agilent Fluorescent Linear Amplification Kit. The fluorescently labeled cRNAs were mixed and hybridized simultaneously to Agilent mouse oligo arrays. The sample pair was hybridized to two arrays, employing a fluor reversal. Chips were scanned with an Agilent dual-laser scanner. Gene expression data were generated using the Agilent feature extraction software (v7.5), with defaults for all parameters.
