Figure 5.

Krüppel‐like factor 2 (KLF2) interacts with Gli1 in hepatocellular carcinoma cells. A, A GST pull‐down assay was used to detect the interaction between recombinant protein GST‐KLF2 and Gli1 expressed by liver cancer cells. B, Co‐immunoprecipitation was used to detect the interaction between KLF2 (Flag‐KLF2) and Gli1 (HA‐Gli1). C, Co‐immunoprecipitation was used to detect the interaction between endogenously expressed KLF2 and Gli1. D, Immunofluorescence staining was used to detect the co‐localization between KLF2 and Gli1 (scale bar: 20 μm, representative images). E, Co‐immunoprecipitation was used to detect the domain of Gli1 for the binding of KLF2. F, Reporter assay was used to examine the effects KLF2 on the activation of Hedgehog/Gli1 signaling by HDAC1. G, Co‐immunoprecipitation was used to detect the effect of exogenously expressed KLF2 (Flag‐KLF2) on the interaction between HDAC1 and Gli1. Transient transfection was performed using the Lipofectamine 2000 according to the manufacturer's instructions