IFI16 Induction by Glucose Restriction in Human Fibroblasts Contributes to Autophagy through Activation of the ATM/AMPK/p53 Pathway
Figure 5
The expression of IFI16 protein is required for the activation of the ATM/AMPK/p53 pathway in response to glucose restriction.
(A) Total RNA isolated from WI-38 HDFs either infected with control or IFI16 shRNA lentivirus particles was subjected cDNA synthesis, followed by quantitative real-time PCR using the TaqMan assay for the IFI16 gene. (B) Sub-confluent cultures of young WI-38 HDFs either infected with control (Vector) or IFI16 shRNA lentivirus particles (shIFI16) were incubated with the indicated concentrations (mM) of glucose in the medium supplemented with 10% fetal bovine serum for 18 h. At the end of the incubations, cells were lysed and equal amounts of proteins were analyzed by immunoblotting using antibodies specific to the indicated proteins. (C) Sub-confluent cultures of young WI-38 HDFs either infected with control (top panel) or IFI16 shRNA lentivirus particles (lower panel) were incubated with the indicated concentrations (mM) of glucose in the medium supplemented with 10% fetal bovine serum for 18 h. At the end of the incubations, cells were photographed using a phase contrast microscope.