Journal: Cellular and Molecular Life Sciences

Article Title: A look into retinal organoids: methods, analytical techniques, and applications

doi: 10.1007/s00018-021-03917-4

Figure Lengend Snippet: A wide range of antibodies is available for IHC of retinal organoids

Article Snippet: PMEL17 , RPE , Mouse , Novus Biologicals , NBP244520 , 1/500 , [ ] .

Techniques: Marker, Clinical Proteomics, Membrane

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: Japanese Journal of Radiology

Article Title: Targeted theranostic nanomedicine using targeted CT-imageable particles that release tebentafusp.

doi: 10.1007/s11604-025-01782-w

Figure Lengend Snippet: Graphical abstract. Two componenents of this theranostics. A : Imaging of malignancy using CT-detectable nanoparticles. B : Targeted immunotherapy using particles that release their contents upon irradiation, consisted of following two sub-components. B-1: induction of HLA-A*02–01-GP100 and B-2:formation of immunological synapse and releasing of perforin/granzymes from CD3 + T-cells

Article Snippet: After heat-mediated antigen retrieval with citrate buffer (pH 6), the samples were incubated with the primary antibody for GP-100 (PMEL/Melanoma gp100 (HMB45) Mouse mAb #38,815, Cell Signaling Technology, Danvers, MA, USA) at a 1/250 dilution, or anti-CD3 + antibody (Anti-Mouse CD3 antibody MAB4841-100, R&D Systems Inc., Minneapolis, MN, USA) at a 1:6400 dilution for 30 min at room temperature.

Techniques: Imaging, Irradiation

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

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: Science immunology

Article Title: The tumor-intrinsic role of the m 6 A reader YTHDF2 in regulating immune evasion

doi: 10.1126/sciimmunol.adl2171

Figure Lengend Snippet: ( A and B ) The tumor growth of WT and Ythdf2 -KO MC38 tumors ( n = 5) and B16-OVA tumors ( n = 6) in C57BL/6 mice. C57BL/6 mice were subcutaneously injected with either 1 × 10 6 WT and Ythdf2 -KO MC38 cells (A) or 1 × 10 6 WT and Ythdf2 -KO B16-OVA cells (B). Tumor sizes were measured every other day starting from the 12th or 14th day after tumor inoculation. ( C and D ) Overall survival of immunocompetent mice subcutaneously implanted with WT or Ythdf2 -KO B16-OVA tumor cells ( n = 14 for WT and n = 15 for KO) (C) or MC38 tumor cells ( n = 8 for WT and n = 10 for KO) (D). ( E and F ) WT or Ythdf2 -KO B16-OVA tumors were transplanted from Rag1 −/− to C57BL/6 mice ( n = 4). Tumor volume (E) and tumor weight (F) were measured on the 14th day after the tumor was transplanted. ( G ) A total of 1 × 10 6 WT or Ythdf2 -KO B16-OVA tumor cells were subcutaneously implanted in C57BL/6 mice pretreated with anti–immunoglobulin G (IgG), anti-NK1.1, anti-CD4, and anti-CD8 antibodies ( n = 5 or 6). Tumor growth was monitored by measuring tumor sizes every other day starting from the eighth day after tumor inoculation. ( H and I ) WT or Ythdf2 -KO MC38-OVA (H) and B16-OVA (I) tumor growth in Rag1 −/− mice that were adoptively transferred with OT1 CD8 + T cells. The mice were subcutaneously injected with either 1 × 10 6 WT or Ythdf2 -KO tumor cells. Tumor sizes were measured every other day starting from the sixth day after tumor inoculation ( n = 5). ( J ) WT or Ythdf2 -KO B16F10 tumor growth in Rag1 −/− mice that were adoptively transferred with gp100-stimulated pmel-1 splenocytes. The mice were subcutaneously injected with either 1 × 10 6 WT or Ythdf2 -KO tumor cells. Tumor sizes were measured every other day starting from the sixth day after tumor inoculation ( n = 5). Data are represented as means ± SD. Statistical analysis was performed using two-way ANOVA with a mixed-effects model with P values adjusted by a Holm-Šídák method for multiple comparisons (A, B, G, H, I, and J), unpaired two-tailed t test (E and F), or Kaplan-Meier survival analysis and log-rank test (C and D). The data presented represent one of two or three independent experiments. * P < 0.05, ** P < 0.01, and **** P < 0.001.

Article Snippet: Splenocytes isolated from pmel-1 T cell receptor transgenic mice were stimulated with gp100 25–33 (5 μg/ml; MedChemExpress) and mouse recombinant IL-2 (100 IU/ml) in RPMI 1640 complete medium at 37°C for 24 hours.

Techniques: Injection, Two Tailed Test