Fig. 3.

Generation of HeLa/Mstn KO cells using CRISPR/Cas9. (A) Schematic diagram of sgRNA targeting the human Mstn exon 2 locus. Protospacer sequences are colored grey. The PAM sequence is framed by a black box. (B) Sequencing of PCR amplification products from Mstn mutant clones confirmed the introduction of insertion-deletion polymorphisms (indels) in exon 2. (C) Immunoblot analysis of Mstn expression in HeLa and HeLa/Mstn KO cells. The antibodies used are indicated on the left. (D) Time-course analysis of viable cell numbers in HeLa and HeLa/Mstn KO cells. Data represent the mean ± standard error of the mean based on three independent experiments, which were analyzed by one-way ANOVA. ^**P < 0.01, ^***P < 0.0001. (E) Proliferation of HeLa and HeLa/Mstn KO cells was determined using CCK8 assays. Data represent the mean ± standard error of the mean based on three independent experiments, which were analyzed by one-way ANOVA. ^**P < 0.01, ^***P < 0.0001.