NR4A1 promotes PDGF-BB-induced cell colony formation in soft agar.

PloS one (2014-10-01)
Glenda Eger, Natalia Papadopoulos, Johan Lennartsson, Carl-Henrik Heldin
RESUMEN

The fibroblast mitogen platelet-derived growth factor -BB (PDGF-BB) induces a transient expression of the orphan nuclear receptor NR4A1 (also named Nur77, TR3 or NGFIB). The aim of the present study was to investigate the pathways through which NR4A1 is induced by PDGF-BB and its functional role. We demonstrate that in PDGF-BB stimulated NIH3T3 cells, the MEK1/2 inhibitor CI-1040 strongly represses NR4A1 expression, whereas Erk5 downregulation delays the expression, but does not block it. Moreover, we report that treatment with the NF-κB inhibitor BAY11-7082 suppresses NR4A1 mRNA and protein expression. The majority of NR4A1 in NIH3T3 was found to be localized in the cytoplasm and only a fraction was translocated to the nucleus after continued PDGF-BB treatment. Silencing NR4A1 slightly increased the proliferation rate of NIH3T3 cells; however, it did not affect the chemotactic or survival abilities conferred by PDGF-BB. Moreover, overexpression of NR4A1 promoted anchorage-independent growth of NIH3T3 cells and the glioblastoma cell lines U-105MG and U-251MG. Thus, whereas NR4A1, induced by PDGF-BB, suppresses cell growth on a solid surface, it increases anchorage-independent growth.

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G 418 disulfate salt, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
G 418 disulfate salt solution, 50 mg/mL in H2O, 0.1 μm filtered, BioReagent, suitable for cell culture
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Z-Leu-Leu-Leu-al, ≥90% (HPLC)
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G 418 disulfate salt, potency: ≥720 μg per mg (dried basis)
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G 418 disulfate salt, suitable for plant cell culture, BioReagent
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Anti-NR4A1 (AB1) antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution

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