Fig. 2: cGAS is essential to regulate proper chromosomal segregations.

a Representative confocal images of mitotic cells 1 h after release from nocodazole-induced mitotic arrest. Cells showing bipolar division, multipolar division, lagging chromosome, and a chromatin bridge (from left to right). b Thirty hours after transfection with sicGAS, mitotic cells were collected by shaking the plate, and they were then treated for 16 h with nocodazole and seeded on a PLL-coated coverslip. After 1 h, cells were fixed with 100% ice-chilled methanol, and ICC was performed. Cells showing the indicated chromosomal segregation errors were calculated as percentages. The results are given as the mean ± SD from three independent experiments (n = 300). ***P < 0.001 as assessed by Student’s t-test. c Co-transfection of sicGAS with pcDNA or MYC-cGAS (non-targeting siRNA) in HeLa cells for 20 h before beginning treatment with nocodazole, which lasted for 16 h. Cells arrested in mitosis were collected and reseeded on PLL-coated coverslips. After 1 h, the percentage of micronucleated cells was quantified using ICC. The results are given as the mean ± SD from three independent experiments (n = 300). ***P < 0.001 by Student’s t-test. d cGAS−/− and wild-type HeLa cells were treated with nocodazole for 16 h. Then, mitotic cells were collected by shaking the plate, and the collected cells were seeded on PLL-coated coverslips for 1 h before ICC was performed to analyze the number of cells showing the indicated chromosomal missegregations. The results are given as the mean ± SD from three independent experiments (n = 300). ***P < 0.001 as assessed by Student’s t-test. e cGAS−/− HeLa cells were transfected with pcDNA or MYC-cGAS and then were treated with nocodazole for 16 h to induce mitotic arrest. After release from nocodazole treatment, mitotic cells were seeded on PLL-coated coverslips for 1 h and were then subjected to ICC to quantify the percentage of cells with chromosomal segregation errors. The results are given as the mean ± SD from three independent experiments (n = 300). *P < 0.05, ***P < 0.001 as assessed by Student’s t-test.