Vitronectin Destroyed Intestinal Epithelial Cell Differentiation through Activation of PDE4-Mediated Ferroptosis in Inflammatory Bowel Disease

Figure 3

VTN regulated ferroptosis through CREB. (a) Internal control pGL4.17 and GPX4 promoter reporter plasmid together with pCDNA 3.0 or CREB plasmid were transfected into HT-29 cells, followed by treated with or without VTN for 48 hr, relative luciferase activities unit were measured and analyzed by two-way analysis of variance (ANOVA) and Dunnett’s multiple comparison test, , , n = 3, data presented as the mean ± s.e.m. (b) ChIP analysis of binding of CREB protein to GPX4 gene promoter in CaCO₂ cells treated as indicated. Student’s t-test, data presented as the mean ± s.e.m, relative luciferase activities unit were measured and analyzed by two-way analysis of variance (ANOVA) and Dunnett’s multiple comparison test, , , n = 3. (c) HT-29 cells were transfected with or without CREB for 24 hr, further incubation with VTN was performed for another 24 hr, the total lysate was collected and subjected from WB to examine indicated protein.

(a)

(b)

(c)