human adar1 p110 isoform antibody  (Proteintech)

Journal: bioRxiv

Article Title: All-trans retinoic acid-mediated ADAR1 degradation synergizes with PD-1 blockade to suppress pancreatic cancer

doi: 10.1101/2024.10.20.619300

Figure Lengend Snippet: (A) Western blots showing the protein levels of ADAR1 and PD-L1 in Capan1, HPAC, BT549 and MB-231 cells treated with the indicated concentrations of ATRA for 24 h. PDAC, pancreatic ductal adenocarcinoma; TNBC, triple-negative breast cancer. (B) Western blots showing the protein levels of Adar1 and Pd-l1 in Panc02 and 4T1 cells treated with the indicated doses of ATRA for 24 h. (C) Western blots showing exogenous MYC-GFP-tagged ADAR1 protein expression in HPAC or MB-231 cells treated with ATRA for 24 h. (D) Western blots showing exogenous FLAG-tagged ADAR1 protein expression in BxPC3 cells treated with ATRA for 24 h. (E) Upper, Western blots showing the protein levels of ADAR1 in HPAC cells treated with ATRA or MG132 alone or in combination for the indicated time. Lower, Quantification of relative ADAR1 expression. (F) Co-immunoprecipitation (Co- IP) blots showing polyubiquitination (Ub) and K48-linked ubiquitination (Ub-K48) of ADAR1 in BxPC3 and Capan1 cells exogenously expressing MYC-GFP-tagged ADAR1 and treated with ATRA. (G) Co-IP blots showing polyubiquitination and Ub-K48 of ADAR1 in HPAC cells treated with ATRA or MG132.

Article Snippet: Human ADAR1 (p150 isoform) antibody was purchased from Bethyl Laboratories, Inc. Human ADAR1 (p110 isoform) antibody and vinculin antibody were purchased from Proteintech.

Techniques: Western Blot, Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay

Journal: bioRxiv

Article Title: All-trans retinoic acid-mediated ADAR1 degradation synergizes with PD-1 blockade to suppress pancreatic cancer

doi: 10.1101/2024.10.20.619300

Figure Lengend Snippet: (A) Schematic representation of the treatment schedule in the Panc02 mouse tumor model. Panc02 tumor-bearing mice were given ATRA or placebo pellets (10 mg per mouse, subcutaneously [s.c.], 21-day release). Administration of anti-PD-1 antibody or IgG control (100 μg per mouse, intraperitoneally [i.p.], 3 times weekly) was started along with ATRA or placebo pellets. Tumor growth, body weight, and mouse survival were measured. (B) Left, representative images showing Panc02 tumors harvested from mice that received the indicated treatments. Scale bar, 1.2 cm. Right, tumor weight. Data represent mean ± SD. (C) Tumor growth curves of individual mice injected with Panc02 tumor cells and subjected to the indicated treatments. (D) Mean tumor growth in mice injected with Panc02 tumor cells and subjected to the indicated treatments. Each dot represents the mean tumor size from 9 mice in each treatment group; whiskers represent SD. (E) Survival curves of mice that received the indicated treatments (9 mice/group). (F) Survival curves of KPC mice that received the indicated treatments (3 mice/group). (G) Western blots showing Adar1 and Pd-l1 protein expression levels in Panc02 cells from mice treated with the indicated regimens. (H) Body weight of tumor-bearing mice treated with the indicated treatments. Data are shown as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, not significant.

Article Snippet: Human ADAR1 (p150 isoform) antibody was purchased from Bethyl Laboratories, Inc. Human ADAR1 (p110 isoform) antibody and vinculin antibody were purchased from Proteintech.

Techniques: Control, Injection, Western Blot, Expressing

Journal: bioRxiv

Article Title: All-trans retinoic acid-mediated ADAR1 degradation synergizes with PD-1 blockade to suppress pancreatic cancer

doi: 10.1101/2024.10.20.619300

Figure Lengend Snippet: (A) Heatmap showing differential marker expression in CD45+ tumor-infiltrating lymphocyte (TIL) clusters identified by analysis of time-of-flight mass cytometry data with viSNE and FlowSOM. DC, dendritic cell; MDSC, myeloid-derived suppressor cell; Treg, regulatory T cell; NK, natural killer cell. (B) Annotation of TIL populations based on differential marker expression as shown in the heatmap (A) and Figure S8. TAM, tumor-associated macrophage. (C) Cellular distribution and clustering as defined by tSNE1 and tSNE2, colored by cell phenotype, for Panc02 tumors subjected to the indicated treatments. Data show all normalized viable single cells; analysis was conducted using viSNE and FlowSOM. (D) Percentage of CD8+ T cells among total CD45+ TILs under the indicated treatments. (E) T-cell cytotoxicity (CCK-8) showing effects of the indicated treatments on pancreatic cancer cell viability. Target Panc28 cancer cells were engineered to express Fcγ receptor 2A fused with the luciferase Luc2 at the C- terminus (Panc28-RL2), and then incubated with CD8+ T cells and the CD3 antibody OKT3 for 3 days. Data represent mean ± SD. P values were calculated by Student t test. (F) T-cell cytotoxicity (CCK-8) assay showing effects of ATRA on pancreatic cancer cell viability. Panc28-RL2 cells overexpressing MYC-GFP-ADAR1 or MYC-GFP (control) were incubated with CD8+ T cells and the CD3 antibody OKT3 for 3 days. Data represent mean ± SD. P values calculated by Student t test.

Article Snippet: Human ADAR1 (p150 isoform) antibody was purchased from Bethyl Laboratories, Inc. Human ADAR1 (p110 isoform) antibody and vinculin antibody were purchased from Proteintech.

Techniques: Marker, Expressing, Mass Cytometry, Derivative Assay, CCK-8 Assay, Luciferase, Incubation, Control

Journal: bioRxiv

Article Title: All-trans retinoic acid-mediated ADAR1 degradation synergizes with PD-1 blockade to suppress pancreatic cancer

doi: 10.1101/2024.10.20.619300

Figure Lengend Snippet: (A) ADAR gene expression profiling by GEPIA2. Red dots: tumor tissues; green dots: normal tissues. The bars represent the median expression of certain tumor type or normal tissue. BRCA, invasive breast carcinoma; ESCA, esophageal carcinoma; HNSC, head and neck squamous cell carcinoma; LGG, lower-grade brain glioma; LIHC, liver hepatocellular carcinoma; PAAD, pancreatic adenocarcinoma; SKCM, skin cutaneous melanoma; STAD, stomach adenocarcinoma; TGCT, testicular germ cell tumor. (B) Kaplan-Meier overall survival curves of 135 pancreatic cancer patients in the cohort with high and low ADAR1 expression determined by immunohistochemical staining. (C) Representative images of the immunohistochemical staining of ADAR1 in tumor tissue from patients in the MDA cohort. Scale bars, 50 μm. (D) Survival curves of breast cancer patients from 2 independent cohorts by ADAR1 expression level. (i) Liu et al. , 2014; (ii) TCGA. (E) Representative images of the immunohistochemical staining of ADAR1 and PD-L1 in tumor tissue from pancreatic cancer patients before treatment with ATRA and nivolumab. Scale bars, 100 μm. (F) Kaplan-Meier overall survival curves for 9 pancreatic cancer patients treated with ATRA 45 mg/m 2 /day plus nivolumab or ATRA ≥60 mg/m 2 /day plus nivolumab. (G) Schematic illustration of the mechanism of switching “cold” (immune-excluded) tumors to “hot” (immune-infiltrated) tumors by using ATRA and anti-PD-1 antibody.

Article Snippet: Human ADAR1 (p150 isoform) antibody was purchased from Bethyl Laboratories, Inc. Human ADAR1 (p110 isoform) antibody and vinculin antibody were purchased from Proteintech.

Techniques: Expressing, Immunohistochemical staining, Staining