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. 2021 Jul 29;12(8):751. doi: 10.1038/s41419-021-04039-2

Fig. 4. POLRMT knockout potently inhibits NSCLC cell proliferation and induces apoptosis activation.

Fig. 4

Single stable primary pNSCLC1 cells, expressing the CRISPR/Cas9-POLRMT-KO construct (“ko-POLRMT”) or the CRISPR/Cas9 control construct (“Cas9-C”), were established, and expressions of POLRMT mRNA and listed proteins were tested by qRT-PCR (A) and western blotting (B) assays. Cells were further cultured for applied time periods, and cell viability, proliferation, migration, and invasion were tested by CCK-8 (C), EdU incorporation (D), “Transwell” (E), and “Matrigel Transwell” (F) assays, respectively, and results were quantified and normalized. The relative caspase-3 activity (G), caspase-PARP cleavages (H), and ssDNA contents (ELISA OD, I) as well as mitochondrial depolarization (by recording JC-1 green monomers intensity, J) and cell apoptosis (by recording TUNEL-positive nuclei ratio, K) were tested as well, and results were quantified and normalized. “pare” stands for parental control cells. Data were presented as mean ± standard deviation (SD, n = 5). *P < 0.05 vs. “Cas9-C” cells. The experiments were repeated five times with similar results obtained. Scale bar = 100 μm (D–F).