Fig. 5: In vitro efficacy of dtEVs carrying siKRASG^G12D or TP53 mRNA.

a Western blot of PANC-1 cells treated with various dtEVs carrying endogenous siKRAS^G12D or scramble siRNA (SCR) for 24 h shows silenced KRAS^G12D expression. b Flow cytometry histogram of DNA distribution in PANC-1 cells treated with CD64^ck_αROR1 dtEVs carrying siKRAS^G12D or 50/50 siKRAS^G12D/TP53 mRNA. Significant apoptosis (black arrow) is seen with cells treated with dtEVs carrying two cargoes. Lower panel shows percentage of cells at G1, S and G2/M stages of the cell cycle in PANC-1 cells treated with various stEVs (Flag, CK) and dtEVs (CK + αROR1) carrying either siKRAS^G12D alone or 50/50 siKRAS^G12D/TP53 mRNA. c Western blot of PANC-1 cells treated with various dtEVs carrying either endogenous TP53 or scramble (SCR) mRNA for 24 h. The p53 and p21 expressions were upregulated in cells treated with dtEVs carrying TP53 mRNA. d Western blot of PANC-1 cells treated with Gemcitabine (GEM) and CD64^ck_αROR1 dtEVs carrying endogenous TP53 mRNA for 24 h shows upregulated p53 and p21, and downregulated BCL-xl expressions. e Western blot of PANC-1 cells treated with GEM and CD64^ck_αROR1 dtEVs carrying SCR, siKRAS^G12D, TP53 mRNA, or 50/50 siKRAS^G12D/TP53 mRNA for 24 h. f Percentage of live, dead and apoptotic (Apop) PANC-1 cells treated with various stEVs (Flag, CK) and dtEVs carrying TP53 mRNA in the presence (+) or absence (−) of GEM. Cells treated with CD64^ck_αROR1 dtEVs and GEM led to high cancer cell death. g The treatment of PANC-1 cells with dtEVs, both with and without GEM, elevated oncogene-induced senescence, as evidenced by increased in SA-β-galactosidase activities. Con is EVs from untreated native MEFs. All EVs were delivered at 1 × 10⁶ EVs/cell. All error bars represent s.e.m. over three independent samples. The siRNA sequences are given in Supplementary Table 3. Scale bar in g is 100 μm. a, c, d, e, g Western blots and staining images are representative of three biological independent experiments.