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. 2019 Sep 16;2(5):e201900358. doi: 10.26508/lsa.201900358

Figure 1. Generation and validation of human TDP-43 KO cells.

Figure 1.

(A, B) Immunoblot analysis of polyclonal populations of HeLa cells transfected with Cas9 plus two sgRNAs against TDP-43 separately or in combination. Ribosomal protein S6 serves as a loading control (B) Immunoblot analysis of a clonal population of TDP-43 KO HeLa cells. (C) Immunoblot evaluation of the restoration of TDP-43 expression via lentiviral transduction of TDP-43 KO cells with an untagged, wild-type form of TDP-43. (D) Immunoblotting revealed that levels of the larger POLDIP3 isoform were rescued by re-expression of TDP-43 in the TDP-43 KO cells. (E) Assessment of POLDIP3 exon 3 inclusion by RT-PCR. (F) Confocal immunofluorescence microscopy shows similar nuclear localization of the TDP-43 protein in both parental HeLa cells and the rescue cell line. Scale bar = 10 μm.