Figure 2
ILF2 interacts with 3Dpol both in vitro and in vivo. A The cells in the HA group were collected for CO-IP assay using anti-HA, and the isolated proteins from the CO-IP assay were then detected by Western blot assay using anti-MYC and anti-HA were used in this assay (1:3000). The cells in the input group were lysed and subjected to Western blot analysis using anti-MYC and anti-HA. B The DEFs cells were transfected with 4 μg of recombinant plasmids pCMV-HA-3D, the expression level of 3D-HA at 24, 48, and 72 hpt were measured by Western blot using anti-HA mAb (1:3000). GAPDH was detected using anti-GAPDH (1:3000). C The cells in the HA group were collected for CO-IP assay using anti-HA mAb, and the isolated proteins from the CO-IP assay were then detected by Western blot assay using anti-ILF2 (1:100) and anti-HA (1:3000). The cells in the input group were lysed and subjected to Western blot analysis using anti-HA mAb (1:3000) and anti-ILF2 (1:100). D The laser confocal microscopy result of the co-localization result of 3D and ILF2 in DEFs at 12 and 24 hpt. The anti-ILF2 (1:50) were used to detect the cellular localization of endogenous ILF2 (red) in Mock DEFs (non-transfected DEFs) and DEFs transfecting with pCMV-HA-3D, while the anti-HA (1:3000) were used to measure the cellular localization of 3D-HA (green). The Mock group indicated blank DEFs, the DAPI channel indicated staining of DEFs nucleus, the Merge images represent single stack of 3D-HA (green), endogenous ILF2 (red), and DAPI (blue). Bar represents 10 μm.