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. 2017 Nov 8;8:1507. doi: 10.3389/fimmu.2017.01507

Figure 7.

Figure 7

Leptin reduces endoplasmic reticulum (ER) stress via the blockade of activating transcription factor 4 (Atf4) transcription in mice adipocytes. (A) Gene expression of Krüppel-like factor 4 (KLF4) in adipocytes incubated with tunicamycin (TM) or leptin (n = 3). (B) Dual luciferase reporter assay of KLF4 and Atf4. Cells were transfected with PGL3-basic or PGL3-Atf4 plasmids, and pc-KLF4 plasmid (n = 3). (C) Chromatin immunoprecipitation (ChIP) analysis between Atf4 and KLF4 (n = 3). (D) Gene expression levels of KLF4, Atf4, Chop, Beclin1, Atg5, IL-18, and IL-1β in adipocytes infected with pAd-KLF4 or si-KLF4 and incubated with leptin or not (n = 3). (E) Graphic representation of a network of the target genes. Bioinformatics analysis of the protein–protein interaction. (F) Atg5 interacted with Atf4. Coimmunoprecipitation (Co-IP) analysis was done in HA-Atg5 and His-Atf4 transfected HEK293 cells. (G) Co-IP analysis was done in pAd-Atg5 and pAd-Atf4 transfected adipocytes with no infection were used as control. Values are means ± SEM. *p < 0.05 compared with the TM group, #p < 0.05 compared with the leptin treatment group.