Journal: International journal of cancer

Article Title: Antitumor immune response of dendritic cells (DCs) expressing tumor-associated antigens derived from induced pluripotent stem cells: in comparison to bone marrow-derived DCs.

doi: 10.1002/ijc.28367

Figure Lengend Snippet: Figure 3. Expression of hgp100 in the genetically modified DCs. The intracellular expression of hgp100 in the genetically modified BMDCs and iPSDCs. DCs were analyzed using intracellular staining flow cytometry. The staining patterns of hgp100 (black) and FITC- matched controls (thin lines) are shown in histograms.

Article Snippet: Intracellular staining with anti-melanoma (hgp100) polyclonal antibodies (Acris Antibodies, Herford, Germany) and FITC-conjugated anti-goat IgG polyclonal antibodies (Vector Laboratories) was performed using a Fixation and Permeabilization Solution Kit (BD Biosciences, San Jose, CA).

Techniques: Expressing, Staining, Cytometry

Journal: International journal of cancer

Article Title: Antitumor immune response of dendritic cells (DCs) expressing tumor-associated antigens derived from induced pluripotent stem cells: in comparison to bone marrow-derived DCs.

doi: 10.1002/ijc.28367

Figure Lengend Snippet: Figure 5. Therapeutic efficacy of genetically modified DCs in the subcutaneous tumor model. Tumor growth suppression in the mice immunized with genetically modified DCs in the subcutaneous tumor model (n 5 5 for each group). The genetically modified DCs were as follows: PBS (———), BMDCs-AxCALacz (—— ), BMDCs- AxCAhgp100 (), iPSDCs-AxCALacZ (– –) and iPSDCs- AxCAhgp100 (——). The results are presented as the mean tumor volume 6 SD of the mice that developed tumors in each group. *Significantly higher therapeutic efficacy than that observed in the other cells (Day 20, p < 0.0001). **No significant differences com- pared to the genetically modified DCs expressing hgp100 in terms of therapeutic efficacy (Day 20, p > 0.05).

Article Snippet: Intracellular staining with anti-melanoma (hgp100) polyclonal antibodies (Acris Antibodies, Herford, Germany) and FITC-conjugated anti-goat IgG polyclonal antibodies (Vector Laboratories) was performed using a Fixation and Permeabilization Solution Kit (BD Biosciences, San Jose, CA).

Techniques: Expressing

Journal: Scientific Reports

Article Title: A model workflow for microfluidic enrichment and genetic analysis of circulating melanoma cells

doi: 10.1038/s41598-025-99153-y

Figure Lengend Snippet: Customized antibody cocktail for live cell staining usingParsortix system.

Article Snippet: GP100 , HMB45 , 1:80 , AF488; 498/520 , Mouse IgG1, Kappa , , Novus Biologicals (NBP2-34638 AF488) , CMC identification.

Techniques: Staining, Concentration Assay

Journal: The Journal of investigative dermatology

Article Title: CXCL5 as Regulator of Neutrophil Function in Cutaneous Melanoma.

doi: 10.1016/j.jid.2018.07.006

Figure Lengend Snippet: Figure 1. Expression of neutrophil-associated genes is increased in CXCL5high melanoma samples of publicly available datasets. (a) Log2FC expression of neutrophil-associated genes in CXCL5high expressing tumors compared with CXCL5low expressing primary melanomas or metastases for five different GEO datasets. (b) Median z-scores of 40 neutrophil-associated genes positively correlate with increasing z-scores of CXCL5 (ranked values) for each individual sample of GEO and TCGA datasets combined (n ¼ 690).

Article Snippet: Mice were killed 16 days later, and the presence of metastasis was evaluated macroscopically (number of black nodules) and microscopically on nonconsecutive histological sections (hematoxylin and eosin and immunohistochemistry with rabbit anti-mouse pmel17/SILV polyclonal antibody [NBP1-69571, Novus Biologicals, Littleton, CO]).

Techniques: Expressing

Journal: Immunity

Article Title: Reactive Neutrophil Responses Dependent on the Receptor Tyrosine Kinase c-MET Limit Cancer Immunotherapy.

doi: 10.1016/j.immuni.2017.09.012

Figure Lengend Snippet: Figure 1. Concomitant Short-Term c-MET Inhibition Enhances the Efficacy of ACT Immunotherapy in Both c-MET-Dependent and c-MET- Independent Melanoma Models (A and B) Left: Effect of the c-MET inhibitor capmatinib on in vitro growth of melanoma cells shown in percent relative to controls (n = 3, mean ± SEM). Right: In vivo growth kinetics of tumor transplants treated as indicated (n = 6; mean tumor area ± SEM). (C and D) Experimental protocols for concomitant capmatinib treatment (5 mg/kg b.i.d.) with ACT immunotherapy (top; C, cyclophosphamide; LV, adenoviral vaccination and Pmel-1 T cell transfer; I, innate immune activation with poly(I:C) and CpG) and Kaplan-Meier survival curves for cohorts of mice treated as indicated (bottom; n = 10 or 15 for HCmel12, n = 10 or 14 for B16F1; numbers behind curves indicate mice with eradicated tumors). (E and F) Representative pictures of vitiligo-like fur depigmentation on back skin (left) and corresponding quantification (right) in individual long-term surviving mice (n = 6 for HCmel12, n = 9 for B16F1). Cumulative results of three independent experiments are shown in (C)–(F). Statistics: unpaired two-tailed Student’s t test *p < 0.05 (A and B); log rank test for survival curves (C and D). See also Figure S1.

Article Snippet: Blots were immunostainedwith p44/42MAPK (ERK1/2) rabbit monoclonal antibody (#9102; Cell Signaling), phospho-ERK (E-4) mousemonoclonal antibody (sc-7383; Santa Cruz) for in vitro samples or phospho-ERK rabbit monoclonal antibody (#4370; Cell Signaling) for tumor samples, AKT (pan) (40D4) mouse monoclonal antibody (#2920; Cell Signaling) for in vitro samples or AKT (pan) (C67E7) rabbit monoclonal antibody (#4691; Cell Signaling) for tumor lysates; phospho-AKT (Ser473) (D9E) rabbit monoclonal antibody (#4060; Cell Signaling), b-Actin (C4) mouse monoclonal antibody (sc-47778; Santa Cruz), c-MET (B-2) mouse monoclonal antibody (sc-8057; Santa Cruz), phospho-MET (Tyr1234/1235) (D26) rabbit monoclonal antibody (#3077; Cell Signaling) and gp100 goat polyclonal antibody (#NB100-41098; Novus Biologicals).

Techniques: Inhibition, In Vitro, In Vivo, Activation Assay, Two Tailed Test

Journal: Immunity

Article Title: Reactive Neutrophil Responses Dependent on the Receptor Tyrosine Kinase c-MET Limit Cancer Immunotherapy.

doi: 10.1016/j.immuni.2017.09.012

Figure Lengend Snippet: Figure 2. Combining c-MET Inhibition with ACT Immunotherapy Increases the Infiltration of Tumors with Pmel-1 T Cells and Decreases the Reactive Recruitment of c-MET+ Neutrophils (A) Experimental protocol for the analysis of immune cells in blood, tumors, and bone marrow. (B) Flow cytometric quantification of Pmel-1 T cells in the blood and tumors of mice treated as indicated (n = 12; mean ± SEM). (C) Representative flow cytometric plots showing Granzyme B (GrzB) expression on CD90.1+CD8+ Pmel-1 T cells in the tumor (left) and quantification in individual mice treated as indicated (right, n = 7; mean ± SEM). (D) Corresponding data for KLRG1 (n = 10 for ACT, n = 9 for ACT+capmatinib; mean ± SEM). (E) Absolute neutrophil counts in the blood of mice on day 22 (for control and capmatinib) or day 28 (for ACT and ACT+capmatinib) treated as indicated (n = 5 for control and capmatinib, n = 12 for ACT and ACT+capmatinib; mean ± SEM). (F) Gating strategy for c-MET+ neutrophils in BMMNCs. (G) Quantification of total and c-MET+ neutrophils in BMMNCs of mice treated as indicated (n = 12; mean ± SEM). (H) Flow cytometric quantification of total and c-MET+ neutrophils in tumors of mice treated as indicated (n = 12; mean ± SEM). Cumulative results of three independent experiments are shown. Statistics: unpaired two-tailed Student’s t test *p < 0.05, **p < 0.01 (E); unpaired two-tailed Mann-Whitney test *p < 0.05, **p < 0.01, and ***p < 0.001 (B–D, G–H). See also Figure S2 and Table S1.

Article Snippet: Blots were immunostainedwith p44/42MAPK (ERK1/2) rabbit monoclonal antibody (#9102; Cell Signaling), phospho-ERK (E-4) mousemonoclonal antibody (sc-7383; Santa Cruz) for in vitro samples or phospho-ERK rabbit monoclonal antibody (#4370; Cell Signaling) for tumor samples, AKT (pan) (40D4) mouse monoclonal antibody (#2920; Cell Signaling) for in vitro samples or AKT (pan) (C67E7) rabbit monoclonal antibody (#4691; Cell Signaling) for tumor lysates; phospho-AKT (Ser473) (D9E) rabbit monoclonal antibody (#4060; Cell Signaling), b-Actin (C4) mouse monoclonal antibody (sc-47778; Santa Cruz), c-MET (B-2) mouse monoclonal antibody (sc-8057; Santa Cruz), phospho-MET (Tyr1234/1235) (D26) rabbit monoclonal antibody (#3077; Cell Signaling) and gp100 goat polyclonal antibody (#NB100-41098; Novus Biologicals).

Techniques: Inhibition, Expressing, Control, Two Tailed Test, MANN-WHITNEY

Journal: Immunity

Article Title: Reactive Neutrophil Responses Dependent on the Receptor Tyrosine Kinase c-MET Limit Cancer Immunotherapy.

doi: 10.1016/j.immuni.2017.09.012

Figure Lengend Snippet: Figure 4. c-MET Inhibition Decreases the Reactive Recruitment of c-MET+ Immunosuppressive Neutrophils in Lymph Node Tissues and Pro- motes Pmel-1 T Cell Expansion (A) Experimental protocol for the analysis of neutrophils and Pmel-1 T cells in the B16F1 melanoma model. (B) Flow cytometric quantification of total and c-MET+ neutrophils in the tumor draining lymph node of mice treated as indicated (right; n = 12; mean ± SEM). (C) Flow cytometric quantification of Pmel-1 T cells (left, n = 12; mean ± SEM) and of Ki67+ expression in Pmel-1 T cells (right, n = 6; mean ± SEM) in the tumor draining lymph node of mice treated as indicated. (D) Experimental protocol for the analyses of neutrophils in tumor draining lymph nodes of Ly6GCreROSA26LSL-tdTomato mice treated with ACT immunotherapy. (E) Representative fluorescent images showing the localization of tdTomato+ neutrophils and adoptively transferred GFP+ Pmel-1 T cells in the tumor draining lymph node on day 10 after ACT immunotherapy. White arrows indicate interaction between neutrophils and Pmel-1 T cells. Cumulative results of three independent experiments are shown. Statistics: unpaired two-tailed Mann-Whitney test *p < 0.05, **p < 0.01, ***p < 0.001 (B and C). See also Figures S3 and S4.

Article Snippet: Blots were immunostainedwith p44/42MAPK (ERK1/2) rabbit monoclonal antibody (#9102; Cell Signaling), phospho-ERK (E-4) mousemonoclonal antibody (sc-7383; Santa Cruz) for in vitro samples or phospho-ERK rabbit monoclonal antibody (#4370; Cell Signaling) for tumor samples, AKT (pan) (40D4) mouse monoclonal antibody (#2920; Cell Signaling) for in vitro samples or AKT (pan) (C67E7) rabbit monoclonal antibody (#4691; Cell Signaling) for tumor lysates; phospho-AKT (Ser473) (D9E) rabbit monoclonal antibody (#4060; Cell Signaling), b-Actin (C4) mouse monoclonal antibody (sc-47778; Santa Cruz), c-MET (B-2) mouse monoclonal antibody (sc-8057; Santa Cruz), phospho-MET (Tyr1234/1235) (D26) rabbit monoclonal antibody (#3077; Cell Signaling) and gp100 goat polyclonal antibody (#NB100-41098; Novus Biologicals).

Techniques: Inhibition, Expressing, Two Tailed Test, MANN-WHITNEY