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. 2021 Nov 8;221(1):e202106100. doi: 10.1083/jcb.202106100

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© 2021 Pennauer et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

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Figure 1. — Characterization of viable Arf KO cell lines. (A) Arf KO HeLaα cell lines were generated by CRISPR/Cas9 as described in Materials and methods and analyzed for partial deletion of exon 1 by genomic PCR. Uniform shortening of PCR fragments in KO cell lines indicated successful genomic deletion in all corresponding Arf alleles. (B) The deletion of Arfs on the protein level was verified by immunoblot analysis. For each cell line, three biological replicates were analyzed on the same gel. Molecular weight markers are indicated in kilodaltons. (C) Growth curves of Arf KO cell lines were derived from live cell counts obtained in three independent experiments (mean ± SD). Cells were seeded at the same density and monitored for 5 consecutive days. (D) Doubling times were calculated from the growth curves displayed in C. Bars, mean ± SD. Unpaired one-way ANOVA versus parental (ns, P > 0.05; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001).