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Fig. 6 | Journal of Biomedical Science

Fig. 6

From: SB203580 increases G-CSF production via a stem-loop destabilizing element in the 3’ untranslated region in macrophages independently of its effect on p38 MAPK activity

Fig. 6

SB203580 induces the luciferase mRNA and activity in RAW264.7 cells transfected with Gp-LUC-3’UTR. (a and b) RAW264.7 cells were co-transfected with reporter plasmid (Gp-LUC or Gp-LUC-3’UTR) and control plasmid (phRLTK), then, 4 h later, were treated with DMSO or 10 μM SB203580 for 20 h. (a) Photinus luciferase mRNA levels were determined by RT-qPCR, normalized to Renilla luciferase mRNA levels, and expressed relative to the value in Gp-LUC-transfected cells treated with DMSO, set as 1. (b) Photinus and Renilla luciferase activities were measured and Photinus luciferase activity normalized to Renilla luciferase activity and expressed relative to the value in the Gp-LUC-transfected cells treated with DMSO, set as 1. (c) p38α MAPK knockdown cells and control knockdown cells (Luc shRNA) were co-transfected with Gp-LUC-3’UTR and phRLTK and treated as described above, then luciferase activities were determined as in (b) and expressed relative to the value in the DMSO-treated Luc controls, set as 1. The values are the mean ± SD for three independent experiments. *p < 0.05 as indicated. n.s., not significant

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