Journal: bioRxiv

Article Title: Mono-ADP-ribosylation-driven immunosuppression and cross-resistance to therapy through cancer cell intrinsic and extrinsic mechanisms

doi: 10.64898/2026.06.01.729331

Figure Lengend Snippet: (A) qRT-PCR analysis of Parp11 mRNA expression in colon adenocarcinoma cell lines (CT26 and MC38) (n=3) and normal mouse colon tissues (n=15). Data are presented as fold change relative to the normal colon tissues group. (B) qRT-PCR analysis of Parp11 mRNA expression in pancreatic ductal adenocarcinoma cell lines (4662, MH6499c4, and MH6419c5) (n=3) and normal mouse pancreas tissues (n=18). Data are presented as fold change relative to the normal pancreas group. (C) qRT-PCR analysis of Parp11 mRNA levels in MC38 and 4662 cells following treatment with adenosine (100 μM for 4 h). Each group contained three biological replicates. Data are presented as fold change relative to the vehicle group. (D) qRT-PCR analysis of Parp11 mRNA levels in MC38 and 4662 cells following treatment with epinephrine (10 μM for 4 h). Each group contained three biological replicates. Data are presented as fold change relative to the vehicle group. (E) qRT-PCR analysis of Parp11 mRNA levels in MC38 and 4662 cells following treatment with GLP-1 ( – ) (100 nM for 4 h). Each group contained three biological replicates. Data are presented as fold change relative to the vehicle group. Relative mRNA levels were normalized to Gapdh. Each dot represents one biological replicate. Data are presented as mean ± SEM (A–E). Statistical significance was determined using two-tailed unpaired Student’s t test (A–E).

Article Snippet: For stimulation experiments, cells were treated with adenosine (100 μM) (Sigma, CAS A4036), epinephrine (10 μM) (selleckchem, S2521), or GLP-1 ( – ) (100 nM) (Phoenix Pharmaceuticals, 028-13) for 4 h unless otherwise indicated.

Techniques: Quantitative RT-PCR, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Mono-ADP-ribosylation-driven immunosuppression and cross-resistance to therapy through cancer cell intrinsic and extrinsic mechanisms

doi: 10.64898/2026.06.01.729331

Figure Lengend Snippet: (A) Flow cytometry analysis of the frequency (% of live CD45 + cells) of CD4 + T cells, CD8 + T cells, and NK1.1 + or CD49b + cells in the spleens from C57BL6 or Balb/c mice bearing subcutaneous tumors described in . (B) Gating strategy for immune profiling of tumor and splenic tissues. (C) Flow cytometry analysis of the frequency (% of live CD45 + cells) and numbers of tumor-associated macrophages (TAMs, CD11b + F4/80 + ), polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs, CD11b + Ly6G + ), and monocytic myeloid-derived suppressor cells (M-MDSCs, CD11b + Ly6C + ), in subcutaneous MC38 or CT26 tumors described in and . (D) Flow cytometry analysis of the frequency and number of CD4 + T cells and NK cells in subcutaneous MC38 or CT26 tumors described in and . (E) Flow cytometry analysis of the percentage of IFN-γ + , Granzyme B + , and perforin + OT-1 T cells in the co-culture system described in (n = 3). (F) Flow cytometry analysis of MFI of H2-KbDb on splenocytes isolated from spleens of WT or Parp11 −/− mice (n = 3). (G) Flow cytometry analysis of MFI of H2-KbDb on MC38 cells pretreated with adenosine (100 μM), epinephrine (10 μM), GLP-1 ( – ) (100 nM), or vehicle for 4 h (n = 3). Data are presented as fold change relative to the vehicle group. Each dot represents one biological replicate. Data are presented as mean ± SEM. Statistical significance was determined using two-tailed unpaired Student’s t test (A, C–G).

Article Snippet: For stimulation experiments, cells were treated with adenosine (100 μM) (Sigma, CAS A4036), epinephrine (10 μM) (selleckchem, S2521), or GLP-1 ( – ) (100 nM) (Phoenix Pharmaceuticals, 028-13) for 4 h unless otherwise indicated.

Techniques: Flow Cytometry, Derivative Assay, Co-Culture Assay, Isolation, Two Tailed Test

Journal: bioRxiv

Article Title: Mono-ADP-ribosylation-driven immunosuppression and cross-resistance to therapy through cancer cell intrinsic and extrinsic mechanisms

doi: 10.64898/2026.06.01.729331

Figure Lengend Snippet: (A) Flow cytometry analysis of the frequency (% of live CD45 + cells) and the number (per gram of tumor tissue) of CD8 + T cells from subcutaneous MC38 or CT26 tumors overexpressing wild-type PARP11, PARP11 HY mutant, or EV control in C57BL6 (n = 5) or Balb/c (n = 7) mice, as described in and . (B) Flow cytometry analysis of the percentage of IFN-γ + and Granzyme B + CD8 + T cells isolated from subcutaneous MC38 or CT26 tumors overexpressing wild-type PARP11, PARP11 HY mutant, or EV control in C57BL6 (n = 5) or Balb/c (n = 7) mice, as described in and . (C) Killing of MC38-OVA-luc cells pre-treated or not with ITK-7 (1 μM, 48 h) or vehicle, followed by adenosine (100 μM), or epinephrine (10 μM), or GLP-1 (100 nM) before co-culture with OT-1 CTLs for 12 h (OT-1: MC38-OVA-luc = 5:1; n = 3). (D) Killing of MC38-OVA-luc cells or B16F10-OVA-luc cells pretreated with ITK-7 (1 μM, 48 h) or vehicle prior to co-culture with OT-1 CTLs for 12 h (OT-1: MC38-OVA-luc = 5:1; OT-1: B16F10-OVA-luc = 5:1; n = 3). (E) Flow cytometry analysis of mean fluorescence intensity (MFI) of H2-Kd on malignant (CD45 − Podoplanin + ) cells from subcutaneous CT26 tumors overexpressing wild-type PARP11, PARP11 HY mutant, or EV control in Balb/c mice (n = 7), as described in . (F) Flow cytometry analysis of MFI of H2-KbDb on malignant (CD45 − EpCAM + ) cells from subcutaneous MC38 tumors overexpressing wild-type PARP11, PARP11 HY mutant, or EV control in C57BL6 mice (n = 5), as described in . (G) Flow cytometry analysis of MFI of H2-KbDb on malignant (CD45 − EpCAM + ) cells from subcutaneous MC38 tumors expressing sgParp11 or NTC in C57BL6 mice (n = 5), as described in . (H) Flow cytometry analysis of MFI of H2-Kd on malignant (CD45 − PDPN + ) cells from subcutaneous CT26 tumors treated as described in (n = 8). Each dot represents one biological replicate. Data are presented as mean ± SEM. Statistical significance was determined using two-tailed unpaired Student’s t test (A–H).

Article Snippet: For stimulation experiments, cells were treated with adenosine (100 μM) (Sigma, CAS A4036), epinephrine (10 μM) (selleckchem, S2521), or GLP-1 ( – ) (100 nM) (Phoenix Pharmaceuticals, 028-13) for 4 h unless otherwise indicated.

Techniques: Flow Cytometry, Mutagenesis, Control, Isolation, Co-Culture Assay, Fluorescence, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Mono-ADP-ribosylation-driven immunosuppression and cross-resistance to therapy through cancer cell intrinsic and extrinsic mechanisms

doi: 10.64898/2026.06.01.729331

Figure Lengend Snippet: (A) Flow cytometry analysis of MFI of H2-KbDb on MC38 cells. Tumor cells were pretreated with ITK-7 (1 μM, 48 h) or DMSO, followed by adenosine (100 μM), epinephrine (10 μM), or GLP-1 (100 nM), or vehicle for 4 h prior to analysis (n = 3). Data are presented as fold change relative to the vehicle group. (B) Box plot showing PARP11 mRNA expression levels in 179 human pancreatic adenocarcinoma (PAAD) samples and 171 normal tissue samples from TCGA. *P < 0.05 was considered statistically significant. (C) Flow cytometry analysis of MFI of H2-KbDb on 4662, MH6499c4, and MH6419c5 cells pretreated with ITK-7 (1 μM, 48 h) or DMSO. Data are presented as fold change relative to the control group. (D) Representative tumor images and tumor weights of subcutaneous MH6419c5 tumors. Mice were treated as described in . Each dot represents one biological replicate. Data are presented as mean ± SEM. Statistical significance was determined using two-tailed unpaired Student’s t test for (A), (C) and (D).

Article Snippet: For stimulation experiments, cells were treated with adenosine (100 μM) (Sigma, CAS A4036), epinephrine (10 μM) (selleckchem, S2521), or GLP-1 ( – ) (100 nM) (Phoenix Pharmaceuticals, 028-13) for 4 h unless otherwise indicated.

Techniques: Flow Cytometry, Expressing, Control, Two Tailed Test