Fig 1: Validation of knock-in FLAG epitope in Nup107. A) Subcellular localization of Flag-tagged Nup107. Exogenously expressed Nup107-3xFLAG was introduced by retrovirus infection. The nuclear membrane was stained with TPR(BETHYL#A300-828A, shown in blue); Nup107-FLAG/3xFLAG was detected by FLAG-M2 antibody (Sigma#F3165, shown in red). Alexa Flour 488 and 568 (Thermo Fisher Sceintific A-32723 and A-11011) were used as fluorophores. Last two columns were zoomed images of the red square portion in the third right panels. B) Validation of the HindIII site insertion by electrophoresis. In knock-in cells, the 1.3 kb PCR product can be digested by HindIII, resulted in one 900 bp and one 400 bp fragment. C) Example PCR scheme for a sequencing based validation.