Figure 3.

C‐terminal tagging of ATP6V1G1. A, ATP6V1G1 was tagged at its C terminus with EmGFP. B, An antisense strand targeting gRNA and PAM site was selected which would create a DSB slightly upstream of the endogenous stop codon. EmGFP was amplified with CHoP‐In primers encoding sense orientation gRNA and PAM sites. C, Good colocalisation of EmGFP signal with the endolysosomal marker Magic Red was seen in a flow cytometry isolated EmGFP positive mixed population (scale bar equals 5 μm). D, Immunoblotting with an antibody against ATP6V1G1 confirmed expression of higher molecular weight, EmGFP‐tagged ATP6V1G1 from its endogenous locus. E, Off target expression of EmGFP was assessed by examining mixed populations of cells for aberrant GFP localisation