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Figure 8 | Molecular Cancer

Figure 8

From: NF-κB induces miR-148a to sustain TGF-β/Smad signaling activation in glioblastoma

Figure 8

NF-κB induces miR-148a expression. (A) Real-time PCR analysis of miR-148a expression in glioblastoma cells treated with 10 ng/mL TNF-α or 100 pM TGF-β for 3 hours. Transcript levels were normalized to U6 expression. (B) Real-time PCR analysis of miR-148a expression in cells treated with or without PDTC or neutralizing anti–NF-κB antibody (2 μg/mL) for 3 hours. (C) Schematic of typical miR-148a promoter SREs. Also shown are ChIP assay results for the miR-148a promoter SREs physically associated with NF-κB. ChIP assay results for the SREs of MIR148a promoter physically associated with NFκB or RNA pol II in indicated cells treated with or without TNF-α (see Supplemental Figure 3). (D) Luciferase assay of cells transfected with the pGL3-SRE reporter treated with or without TNF-α. (E) Smad luciferase reporter activity in cells treated with or without TNF-α for 3 hours (F) WB of p-Smad2, p-Smad3, Smad2/3, and IκBα in TNF-α–treated cells (10 ng/mL) in response to treatment with control. (G) In vitro kinase assay indicating that TGF-β–induced endogenous Smad activity was prolonged in TNF-α–treated cells, which the miR-148a inhibitor abrogated. Error bars represent the mean ± SD of three independent experiments. *P < 0.05.

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