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Image Search Results
Journal: Molecular and Cellular Biology
Article Title: DCNL1 Functions as a Substrate Sensor and Activator of Cullin 2-RING Ligase
doi: 10.1128/MCB.01342-12
Figure Lengend Snippet: DCNL1 modulates HIF1α activity. (A) HEK293 cells were transfected with nontargeting scrambled siRNA (siSCR) or DCNL1-targeting siRNA (siDCNL1). An equivalent amount of protein lysate was resolved by SDS-PAGE and detected by immunoblotting using the indicated antibodies. (B) Stable cell lines in the indicated cellular backgrounds were generated through lentivirus-mediated infection of two different shRNA constructs targeting DCNL1. Cells were treated with MG132 for 4 h and lysed, and an equivalent amount of protein lysate was resolved by SDS-PAGE. The indicated proteins were detected by immunoblotting. (C) The indicated stable cell lines were transiently transfected with plasmids encoding firefly luciferase under the control of a hypoxia response element and a Renilla luciferase control. Cells were treated with MG132 for 4 h prior to a dual-luciferase assay. pGIPZ was arbitrarily set to 100 to represent normal HIF1α activity. Error bars represent standard deviations of the mean. An unpaired t test was performed to assess the statistical significance.
Article Snippet: Cells were transfected in triplicate with pGL3-5×HRE-Luc (where Luc is luciferase) (kind gift from Richard P. Hill) and pRL-SV40 (where SV40 is simian virus 40) (
Techniques: Activity Assay, Transfection, SDS Page, Western Blot, Stable Transfection, Generated, Infection, shRNA, Construct, Luciferase, Control