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Fig. 4 | Journal of Neuroinflammation

Fig. 4

From: CCAAT/enhancer-binding protein delta regulates the stemness of glioma stem-like cells through activating PDGFA expression upon inflammatory stimulation

Fig. 4

CEBPD regulates PDGFA transcription through PDGFA promoter region. a The PDGFA mRNA levels in U373MG spheroid cells stably expressing shLacZ or shC7 cells. Real-time PCR analyses conducted with specific primers using cDNA harvested from 5 ng/mL IL-1β treatment on day 3 and 6. Cells transduced with shLacZ served as the sham control. b The identification of CEBPD binding motifs on the PDGFA promoter region. The representation of reporter construct is at the upper region. The reporter vector carrying the PDGFA promoter was co-transfected with the indicated expression vectors in U373MG cells. After 12 h, the cell lysates were harvested for Western blot analysis and luciferase assay. Expression of α-tubulin served as the internal control. c CEBPD binding to the PDGFA promoter in vivo. Chromatin from U373MG spheroid cells treated with or without 5 ng/mL IL-1β was isolated, and chromatin immunoprecipitation assays were performed with indicated antibodies. The precipitated DNA was amplified using specific primers of the PDGFA promoter region. (Total data were obtained from three independent experiments, and the data shown here were from one representative assay. All data are expressed as mean ± SEM. Differences among groups were determined with one-way ANOVA followed by Tukey’s multiple comparison test (a) or unpaired two-tailed t test (b). ***p < 0.001) shLacZ, shβ-galactosidase; shC7, shCEBPD

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