Fig. 3

CEBPD regulates MMP8 transcription in human primary astrocytes through the MMP8 promoter region. MMP8 transcription was increased in (A) CEBPD overexpression or (B) ouabain treatment in astrocytes. Q-PCR analyses were performed using specific primers with cDNA harvested from either CEBPD overexpression or 20 μM ouabain treatment in astrocytes. Quantitative data is presented as means ± SEM; N = 3 (A) or N = 4 (B); analyzed using a two-tailed unpaired Student’s t-test, *p < 0.05 and ****p < 0.0001. The protein expression of MMP8 was also increased in (C) CEBPD overexpression or (E) ouabain treatment in astrocytes. Quantitative data from (D) and (F) depict the expression levels of CEBPD in astrocytes. The CEBPD was normalized using β-Actin. Quantitative data is presented as means ± SEM; N = 3; analyzed using a two-tailed unpaired Student’s t-test, **p < 0.01 and ***p < 0.001. G Identification of CEBPD binding motifs on the MMP8 promoter region involved co-transfecting the MMP8 promoter reporter vectors with either the HA or HA-CEBPD expression vectors into human primary astrocytes. After 24 h, cell lysates were harvested for Western blot analysis and luciferase assays. H The schematic illustrates CEBPD-regulated MMP8 expression in astrocytes