Phosphatidylinositol 3-Kinase Regulates Bone Morphogenetic Protein-2 (BMP-2)-induced Myocyte Enhancer Factor 2A-dependent Transcription of BMP-2 Gene in Cardiomyocyte Precursor Cells

Concerted Action of Smad and CREB-binding Protein Regulates Bone Morphogenetic Protein-2-stimulated Osteoblastic Colony-stimulating Factor-1 Expression

“…DNaseI Footprint Assay-Nuclear extract was prepared from C2C12 and fetal rat calvarial osteoblast cells as described earlier following the protocol specified by the vendor (40). The DNA fragment spanning a region, Ϫ627 to Ϫ509 bp, of CSF-1 promoter was cloned into plasmid vector.…”

“…The mutant oligonucleotides were designed by (a) mutating the core binding sequence ATC TGC to gga TtC (MT), (b) mutating DNA sequences 3Ј to the core binding site CGG GTT to att cGg (3Ј-MT), and (c) mutating 5Ј sequence to the core binding site GGG AGC to ata Atg (5Ј-MT). The double-stranded oligonucleotides were labeled using [␥- 32 P]ATP and T4 polynucleotide kinase (40). The EMSA was performed using 5 g of nuclear extract as described previously (40 -42).…”

Bone Morphogenetic Protein 9 and 13 Induce C3H10T1/2 Cell Differentiation to Cardiomyocyte-Like Cells in Vitro

“…These data suggested that BMP9 might be a better one to induce cardiac differentiation of C3H10T1/2 cells over BMP13. Previous studies have shown that overexpression of BMP2 could regulate the expression of Gata4, Mef2c, and Nkx2.5 and induce the differentiation of stem cells into cardiomyocytes through p38MAPK, Smads, and PI-3K signaling pathways (5,9,31,33). Expression of BMP2, VEGF, and FGF was increased after myocardial infarction and might contribute to cell differentiation and angiogenesis (1,8).…”

A Linear Signal Transduction Pathway Involving Phosphatidylinositol 3-Kinase, Protein Kinase Cϵ, and MAPK in Mesangial Cells Regulates Interferon-γ-induced STAT1α Transcriptional Activation

“…The EMSA was performed with the cell lysate essentially as described before (19,(32)(33)(34). For supershift analysis, the cell lysates were incubated with the STAT1␣ antibody before incubation with the probe (19,33,34). The protein-DNA complex was separated by 5% polyacrylamide gel electrophoresis.…”

“…The 15-bp double stranded oligonucleotide probe (SIE) was labeled with [␥-32 P]ATP and T4 polynucleotide kinase as described before (32). The EMSA was performed with the cell lysate essentially as described before (19,(32)(33)(34). For supershift analysis, the cell lysates were incubated with the STAT1␣ antibody before incubation with the probe (19,33,34).…”