Figure 1. Macropinocytosis is not responsible for cryptotanshinone-induced differential sensitization of PDAC cells. (A) CCK-8 assay assessing cell viability after treatment with JTC801, cryptotanshinone (Crypto), or their combination (Drug concentrations: JTC801, 3.5 μM; Crypto, 5 μM; treatment time: 24 h); (B) Cell death assay evaluating the proportion of dead cells under the indicated treatments (Same concentrations and treatment time as A); (C) Western blot analysis of protein expression in the indicated groups (Drug concentration: JTC801, 3.5 μM; treatment time: 24 h); (D) TRITC-dextran uptake to assess macropinocytosis in the indicated cell lines (Drug concentration: JTC801, 3.5 μM; treatment time: 24 h). Scale bar = 50 µm; (E) CCK-8 assay assessing cell viability after treatment with JTC801, Crypto, or Crypto combined with 5% BSA; (F) CCK-8 assay evaluating cell viability after treatment with JTC801, Crypto, or Crypto combined with EIPA (Drug concentrations: JTC801, 3.5 μM; Crypto, 5 μM; EIPA, 5 μM; treatment time: 24 h); (G and H) CCK-8 assay assessing the effect of LY2090314 combined with JTC801 (G) or Crypto (H) on cell viability (Drug concentrations: JTC801, 3.5 μM; Crypto, 5 μM; LY2090314, 5 μM; treatment time: 24 h); (I) CCK-8 assay analyzing MIAPaCa2 cell viability treated with Crypto (5 μM) with ATP6V0D1 knockout alone or simultaneous GSK3B knockout. Data (A and B, D-I) are presented as mean ± SD from ≥ 3 biologically independent samples (A, n = 6; B = 4; D, n = 3; E-I, n = 6). Statistical significance was determined using two-way ANOVA with Tukey’s multiple comparisons test. Western blot data (C) represent three independent experiments. Source data are provided. *P < 0.05; ns: not significant. PDAC: Pancreatic ductal adenocarcinoma; ATP6V0D1: ATPase H+ transporting V0 subunit D1; SD: standard deviation; ANOVA: analysis of variance.
