Thao D. Pham

Supplemental Fig. S6. KPC-FIO cells display reduced sensitivity to various combinations of 5FU (F), SN38 (I), and oxaliplatin (O). A, B. Parental KPC (2138 and 3886) and KPC-FIO (2138-FIO and 3886-FIO) cells were treated with vehicle control, increasing doses of F (0 µM, 0.5 µM, 1 µM, 2 µM), increasing doses of SN38 (0 nM, 50 nM, 100, 200 nM), increasing doses of O (0 µM, 1 µM, 4 µM, 8 µM) or a combination of these doses. After 72 hours, the effects of drug treatments on cell proliferation were evaluated by WST1 assay. Values were normalized against the vehicle-treated sample and visualized by the depicted heatmap. Heatmaps are representative of three biological replicates.

Supplemental Fig. S1. Gating strategy to determine the frequency of T cells (CD45+CD4+ or CD45+CD8+), T cell functions (TNFα, Perforin1 (Prf1), IFNγ, Granzyme B (GzmB)), the frequency of tumor-associated macrophages (TAMs; CD45+CD11b+F4/80+), granulocytic myeloid derived suppressor cells (G-MDSCs; CD45+CD11b+Ly6G + Ly6C-), monocytic myeloid derived suppressor cells (M-MDSCs; CD45+CD11b+Ly6G-Ly6G+) and their functions (CD206, PD-L1, Arginase1 (Arg1)). All cells are gated from live single cells.

Effect of MRTX1133 on ‘small’ KPC tumors.

Venetoclax enhances the efficacy of MRTX1133 in 2138 and 1245 tumors in vivo.

Supplemental Fig. S3. KPC-FIO cells display reduced sensitivity to 5FU. A-D. The parental KPC and KPC-FIO cells were treated with increasing doses of 5FU (F; 2, 4, 8, 16 µM). After 72 hours, the effects of drug treatments on cell proliferation were evaluated by WST1 assay and analyzed by GraphPad. Error bars ± SEM from 3 biological replicates. One-way ANOVA, followed by Dunnett multiple comparison test. *, p<0.05 **, p<0.01 ***, p<0.001 ****, p<0.0001 ns, non-significant.

Supplemental Fig. S7. Trametinib is the most effective MEK inhibitor at blocking ERK phosphorylation in the KPC and KPC-FIO cells. A-C. 2138 and 2138-FIO (A), 3886 and 3886-FIO (B), and 3213 and 3213-FIO (C) cells were treated with vehicle control (-, DMSO), U0126 (2 µM, U), Binimetinib (2 µM, B), Cobimetinib (2 µM, C), Selumetinib (2 µM, S), or Trametinib (2 µM, T). After 2 and 24 hours, the cells were collected and analyzed for p-ERK1/2 and total ERK1/2 by western blotting with HSP90 as loading control. Blots are representative of three independent replicates.

2138-K and 3213-K cells grow rapidly in vivo.

PDAC cells in ‘high’ confluent collagen cultures exhibit reduced proliferation.