Skip to main content
. 2022 May 18;8(20):eabn7382. doi: 10.1126/sciadv.abn7382

Fig. 5. Therapeutic effect and antitumor mechanism in vitro.

Fig. 5.

(A) Cell viability of U87-EGFRvIII cells after incubation with RNSs, RNCOs, RNSs-D, nRNCOs-D, and RNCOs-D. The concentration of DOX (1, 2, 4, 6, and 8 μM) and NCs (0.5, 1, 2, 3, and 4 μM) increased as the RNS concentration increased. Data are means ± SD (n = 3). Statistical significance was determined by unpaired two-tailed Student’s t test (***P < 0.001). (B) Western blot (WB) analysis of apoptosis protein levels in U87-EGFRvIII cells. (C) CLSM of U87-EGFRvIII cells costained with calcein AM/PI after treatment with PBS, DOX, rRNCOs-D, RNSs-D, nRNCOs-D, and RNCOs-D. Scale bar, 100 μm. (D) Cell apoptosis and necrosis analyzed by annexin V–APC/DAPI (4′,6-diamidino-2-phenylindole) staining after treatment of PBS, RNSs, RNCOs, RNSs-D, nRNCOs-D, and RNCOs-D, respectively (from left to right). (E) Schematic illustration of antitumor mechanism. (F) WB analysis of EGFRvIII protein levels. RNCOs-1 refers to 500 nM equivalent Cas13a RNP; RNCOs-2 refers to 1 μM equivalent Cas13a RNP. (G) RNA denaturing gel electrophoresis examining total RNA integrity.