Mechanism of KLF4 Protection against Acute Liver Injury via Inhibition of Apelin Signaling

Figure 4

TNF-α decreases apelin mRNA and protein levels, but increases KLF4 expression in LX-2 cells. All experiments were performed in triplicate. (a, b) LX-2 cells were treated with TNF-α (10 ng/mL) for various times or at different doses (for 24 h). Total RNA was transcribed with reverse transcriptase and amplified by PCR. GAPDH was used as an internal control. (c, d) Western blotting was performed using anti-apelin and anti-KLF4 antibodies to examine protein expression at various times or at different doses. β-Actin was used as a loading control. (e) LX-2 cells were treated with TNF-α (10 ng/mL) for 24 h and the cells were then fixed and stained by immunofluorescence. Expression of apelin was detected in the cytoplasm (magnification: ×200). (f) LX-2 cells were treated with TNF-α (10 ng/mL) for 24 h and then the cells were then fixed and stained by immunofluorescence. The expression of KLF4 was detected in nuclei (magnification: ×200).

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