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. 2017 Apr 5;8(22):36469–36483. doi: 10.18632/oncotarget.16863

Figure 4. Kifc1 knockout also results in the disorganization and dispersal of the Golgi apparatus.

Figure 4

(A) Representative sequences of the human kifc1 locus targeted by Cas9n and the selected sequences showing representative indels. Generation of indels in the human kifc1 locus in HEK293T cells by CRISPR-Cas9 system. Two different kifc1 knockout cell lines were selected as candidates in this study (designated as kifc1−/− clone 1 and clone 2). (B) Western blot analysis of KIFC1 expression in control, kifc1−/− clone 1 and clone 2 HEK293T cells. β-Actin was used as the loading control. (C) Representative confocal images of the Golgi marker GM130 and KIFC1 in control and kifc1 knockout HEK293T cells (kifc1−/− clone 1 and clone 2). DAPI, blue; GM130, green; KIFC1, red. DIC, differential interference contrast. Scale bars: 5 μm. (D) Quantification of the ratios of disorganized Golgi apparatus in control and kifc1 knockout HEK293T cells (n = 3 cultures for each phenotype). Data are presented as means ± SEM. ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001 (Student's t-test). (E) Quantification of the ratios of the dispersal Golgi in control and kifc1 knockout HEK293T cells (n = 3 cultures for each phenotype). Data are presented as means ± SEM. ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001 (Student's t-test). (F) Representative three dimensional images of the Golgi apparatus (DAPI, blue; GM130, green) in the control, kifc1−/− clone 1 and clone 2 HEK293T cells.