Fig. 2.

Expression of high-molecular weight TDP-43 protein verifies successful CRISPR/Cas9-mediated gene insertion of mAvicFP1 and/or mCherry to TARDBP alleles. Six monoclonal cell lines of varying intensity were selected for validation of mAvicFP1, mCherry, or dual-tag insertion. a Immunoblotting for TDP-43 and myc revealed the abundance of ‘untagged’ (43 kDa) and ‘tagged’ TDP-43 (72–74 kDa), which were quantified by densitometry analysis. CRISPR-editing ratios for b TARDBP-mAvicFP1, c TARDBP-mCherry, or d TARDBP-mAvicFP1/-mCherry monoclonal cell lines were calculated by dividing the total protein-normalised signals of high-molecular weight tagged TDP-43 by the combined signals of all TDP-43 bands, as described in Supplementary Fig. S2. ‘#’ Represents a lower-molecular-weight band positive for endogenous mCherry-tagged TDP-43. e Representative confocal images captured at 63 × magnification depict relative fluorescence intensities of TARDBP-tagged cell lines and clone names are indicated in the upper left corner. Note that TARDBP-mAv/mCh images are displayed as a merge of separate mAvicFP1 and mCherry channels (scale = 20 µm)