Gene Expression Profiling of NFATc1-Knockdown in RAW 264.7 Cells: An Alternative Pathway for Macrophage Differentiation

• Published in: Cells (Basel, Switzerland) Vol. 8; no. 2; p. 131
• Main Authors: Russo, Roberta, Mallia, Selene, Zito, Francesca, Lampiasi, Nadia
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 07-02-2019; MDPI
• Subjects: Animals; Antibiotics; Cell differentiation; Cell Differentiation - genetics; Cytokines; GATA2; GATA2 Transcription Factor - metabolism; Gene expression; Gene Expression Profiling; Gene Knockdown Techniques; Gene Ontology; Infrared imaging systems; Macrophages; Macrophages - cytology; Macrophages - metabolism; Mice; Molecular modelling; NFATC Transcription Factors - metabolism; NFATc1-knockdown; Osteoclastogenesis; Osteoclasts; Osteoclasts - cytology; Osteoclasts - metabolism; Osteogenesis; Phosphatase; Protein Interaction Maps; Proteins; RAW 264.7 Cells; STAT6; Stat6 protein; Transcription factors; St Louis Missouri; United States > US; Temecula California; Germany; NFATc1-knockdown; macrophages; STAT6; GATA2; transcription factors; osteoclasts
• ISSN: 2073-4409; 2073-4409
• DOI: 10.3390/cells8020131

NFATc1, which is ubiquitous in many cell types, is the master regulator of osteoclastogenesis. However, the molecular mechanisms by which NFATc1 drives its transcriptional program to produce osteoclasts from macrophages (M) remains poorly understood. We performed quantitative PCR (QPCR) arrays and bioinformatic analyses to discover new direct and indirect NFATc1 targets. The results revealed that NFATc1 significantly modified the expression of 55 genes in untransfected cells and 31 genes after NFATc1-knockdown (≥2). Among them, we focused on 19 common genes that showed changes in the PCR arrays between the two groups of cells. Gene Ontology (GO) demonstrated that genes related to cell differentiation and the development process were significantly (
> 0.05) affected by NFATc1-knockdown. Among all the genes analyzed, we focused on GATA2, which was up-regulated in NFATc1-knockdown cells, while its expression was reduced after NFATc1 rescue. Thus, we suggest GATA2 as a new target of NFATc1. Ingenuity Pathway Analysis (IPA) identified up-regulated GATA2 and the STAT family members as principal nodes involved in cell differentiation. Mechanistically, we demonstrated that STAT6 was activated in parallel with GATA2 in NFATc1-knockdown cells. We suggest an alternative pathway for macrophage differentiation in the absence of NFATc1 due to the GATA2 transcription factor.