Fig 1.

Analysis of COG 7 KO: A) Lectin staining of WT HEK293 and COG 7 KO HEK293 cells with Galanthus nivalis lectin-Alexa 647 (GNL). Cells were mounted in DAPI containing media to visual the nuclei (blue). GNL is false colored pink. Images were takes with a 63x immersion lens on a confocal microscope. Settings were calibrated using stained control cells and kept the same throughout.

B) Western immunoblot for WT HEK and COG 7 KO cells: delta 7 cell line G3 blotted next to WT HEK293 lysate as a control.

C) Sequencing and BLAST of COG gene in KO cells: PCR products were obtained by amplifying the gene of interest from chromosomal DNA. These products were then cleaned and sent for sequencing. Products appeared to be of the expected size by gel (531 bp). Once the sequence was obtained this was BLASTed against the human genome. The expected cut site based on the guide RNA is highlighted in yellow and resultant frameshift mutations can be seen in red. WT HEK293 cells were also sequenced and BLASTed as a control. Frameshift mutations were not seen in control samples.