Journal: Journal for Immunotherapy of Cancer

Article Title: Overcoming immunotherapy resistance in bladder cancer with a novel antibody-drug conjugate RC48

doi: 10.1136/jitc-2025-011881

Figure Lengend Snippet: RC48-ADC therapy reshapes the tumor immune microenvironment in BCa. ( A ) Kaplan-Meier survival curves were generated for the anti-PD-1 cohort, stratifying patients based on ERBB2 expression levels. Kaplan-Meier plotter ( https://kmplot.com/analysis/ ) was used to generate survival curves and assess the efficacy of immunotherapy. (B) Boxplot illustrating the relationship between ERBB2 expression and immunotherapy efficacy in the Xiangya Immunotherapy Cohort. (C) The relationship between ERBB2 expression and immune scores in the TCGA-BLCA cohort. (D) Activity levels of the cancer immunity cycle across high and low ERBB2 expression groups. (E) Correlations between ERBB2 and various immune cell types (CD8+T cells, CD4+T cells, dendritic cells, and natural killer cells) analyzed using five independent algorithms (TIMER, CIBERSORT, CIBERSORT-ABS, MCP-COUNTER, and XCELL). (F) Correlations between ERBB2 and tumor-infiltrating immune cells (TIICs) (right) and the cancer immunity cycle (left). (G) Expression patterns of immune cell-related effector genes (including NK cells, Th1 cells, macrophages, CD8+T cells, and dendritic cells) in ADC-treated (n=7) and ADC-untreated (n=18) groups. (H) Cancer immunity cycle activity in the ADC-treated (n=7) and ADC-untreated (n=18) groups. *p<0.05; **p<0.01; ***p<0.001. (I) Correlations between ERBB2 and various immune cell types. *p<0.05; **p<0.01; ***p<0.001. (J) tSNE plot showing the distribution of immune cells in BCa samples from RC48- untreated and RC48-treated patients, with each dot representing a single cell and colors indicating different cell types. (K) Differences in the ratios of three CD8+T cell subgroups (exhausted CD8+T cells, cytotoxic CD8+T cells, and TRM CD8 cells) estimated using the STARTRAC-dist index in ADC-treated (n=4) and ADC-untreated (n=2) groups. +++, Ro/e>1; ++, 0.8

Article Snippet: The primary antibodies included: Anti-PD-L1 antibody ( EPR19759 ), abcam, ab213524; anti-PD-L1 antibody ( EPR20529 ), abcam, ab213480; anti-CD8 alpha antibody ( EPR21769 ), abcam, ab217344; anti-CD8 alpha antibody (CAL66), abcam, ab237709; Granzyme B Polyclonal antibody, Proteintech, 13588-1-AP; TAZ Rabbit Polyclonal Antibody, Proteintech, 23306-1-AP.

Techniques: Generated, Expressing, Activity Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Overcoming immunotherapy resistance in bladder cancer with a novel antibody-drug conjugate RC48

doi: 10.1136/jitc-2025-011881

Figure Lengend Snippet: RC48-ADC inhibits tumor PD-L1 expression, increases CD8+T cell infiltration and activity. ( A ) h-HER2-MB49 mouse bladder cancer cells overexpressing h-HER2 were injected into mice on day 0, and RC48-ADC was administered at either a high dose (HD, 10 mg/kg) or low dose (LD, 5 mg/kg) based on the indicated schedule. (B) Tumor volumes were measured at various time points, with data shown as mean±SD. (C) Representative tumor images from different RC48-ADC dose groups at the final time point in the h-HER2-MB49 tumor-bearing mouse model. (D) Tumor weights were measured on day 15 following RC48-ADC treatment. (E) Representative flow cytometry profiles showing CD8 (CTL marker) detection in h-HER2-MB49 tumors from different treatment groups. (F) Quantification of CD8+/CD3+ cells in tumors from various treatment groups (n=7 per group). (G) Representative flow cytometry profiles displaying GZMB and IFNγ, markers of T cell activity, in h-HER2-MB49 tumor tissues across different treatment groups. (H, I) Quantification of CD8+GZMB+/IFNγ+ CTL percentages in tumor tissues from different treatment groups (n=7 per group). (J) Representative flow cytometry profiles of PD-L1 detection in h-HER2-MB49 tumor tissues from the various treatment groups. (K) Quantification of PD-L1+CD45 cells in tumors from different treatment groups (n=7 mice per group). (L) Representative images of immunohistochemistry staining for CD8 and PD-L1 in h-HER2-MB49 tumors. (M) Representative immunofluorescence staining images showing CD8 and PD-L1 in h-HER2-MB49 tumors. (N, O) Quantification of CD8+ and PD-L1+cell percentages in tumors from different treatment groups (n=7 per group). (P) Representative images of immunofluorescence staining for CD8, GZMB, and PD-L1 in tumor specimens before and after RC48-ADC treatment. (Q) Quantification of CD8+, GZMB+, and PD-L1+cells in tumors RC48-ADC treated or untreated. (n=13 per group). Data are presented as mean±SD, *p<0.05, **p<0.01, ***p<0.001. ADC, antibody-drug conjugate.

Article Snippet: The primary antibodies included: Anti-PD-L1 antibody ( EPR19759 ), abcam, ab213524; anti-PD-L1 antibody ( EPR20529 ), abcam, ab213480; anti-CD8 alpha antibody ( EPR21769 ), abcam, ab217344; anti-CD8 alpha antibody (CAL66), abcam, ab237709; Granzyme B Polyclonal antibody, Proteintech, 13588-1-AP; TAZ Rabbit Polyclonal Antibody, Proteintech, 23306-1-AP.

Techniques: Expressing, Activity Assay, Injection, Flow Cytometry, Marker, Immunohistochemistry, Staining, Immunofluorescence

Journal: Journal for Immunotherapy of Cancer

Article Title: Overcoming immunotherapy resistance in bladder cancer with a novel antibody-drug conjugate RC48

doi: 10.1136/jitc-2025-011881

Figure Lengend Snippet: RC48-ADC promotes the recruitment and activation of CD8+T cells by inducing the release of chemokines from tumors. ( A) h-HER2-MB49 cells were injected into mice on day 0, and treatment with RC48-ADC (10 mg/kg) and CD8α (100 µg/mouse) was administered based on the indicated schedule. (B) Tumor volume was measured at various time points. Data are presented as mean±SD. (C) Mice were sacrificed on day 15 following RC48-ADC or CD8α treatment, and tumor weights were measured. (D) Representative images of tumors at the final time point after RC48-ADC or CD8α treatment in the h-HER2-MB49 tumor-bearing mouse model. (E) Peripheral blood T cells were extracted and purified for T cell-mediated cytotoxicity assay and Chemotaxis assay. (F) Flow cytometry analysis showing the purification of CD8+T cells for chemotaxis experiments. (G, H) T24 cells were co-incubated with activated T cells for 48 hours, either with or without RC48-ADC (5 µmol), and then stained using crystal violet. The numbers represent the normalized cancer cell survival rates after T cell killing. The ratio of T cells to tumor cells was maintained at 3:1. (I) Bar graph showing the quantitative analysis of cancer cell survival rates from ( E ). (J, K) Flow cytometry analysis showing differences in CD8+T cell activity among various co-culture groups in the T24 or 5637 cell lines. (L-M) Chemotaxis assay demonstrating the different chemotaxis abilities of CTLs in the control versus RC48-ADC-treated group. (N) Bar graph showing the mRNA levels of four chemokines as determined by qRT-PCR after RC48-ADC treatment. (O) Bar graph showing normalized protein secretion concentrations of the four chemokines after RC48-ADC treatment. (P, Q) Use three different neutralizing antibodies to block the chemokines and assess the number of CD8+T cells in T24 and 5637. Data are presented as mean±SD, *p<0.05, **p<0.01, ***p<0.001. ADC, antibody-drug conjugate. FSC, Forward Scatter; SSC, Side Scatter; PBS, phosphate buffered saline.

Article Snippet: The primary antibodies included: Anti-PD-L1 antibody ( EPR19759 ), abcam, ab213524; anti-PD-L1 antibody ( EPR20529 ), abcam, ab213480; anti-CD8 alpha antibody ( EPR21769 ), abcam, ab217344; anti-CD8 alpha antibody (CAL66), abcam, ab237709; Granzyme B Polyclonal antibody, Proteintech, 13588-1-AP; TAZ Rabbit Polyclonal Antibody, Proteintech, 23306-1-AP.

Techniques: Activation Assay, Injection, Purification, Cytotoxicity Assay, Chemotaxis Assay, Flow Cytometry, Incubation, Staining, Activity Assay, Co-Culture Assay, Control, Quantitative RT-PCR, Blocking Assay, Saline

Journal: Journal for Immunotherapy of Cancer

Article Title: Overcoming immunotherapy resistance in bladder cancer with a novel antibody-drug conjugate RC48

doi: 10.1136/jitc-2025-011881

Figure Lengend Snippet: The combination of RC48-ADC and CTLA-4/PD-1 mAb has a synergistic effect in treating BCa in immunocompetent mice. ( A ) h-HER2-MB49 cells were injected into mice on day 0, and treatment with RC48-ADC (10 mg/kg) and 100 µg/mouse of PD-1 or CTLA-4 mAb was administered as indicated. (B) Tumor volume was measured at various time points. (C) Mice were sacrificed on day 15 after treatment with RC48-ADC, PD-1, or CTLA-4 mAb, and tumor weight was measured. (D) Representative tumor images at the end of the experiment after RC48-ADC, PD-1, or CTLA-4 mAb treatment in the h-HER2-MB49 tumor-bearing mouse model. (E) Immunofluorescence staining of CD8 and PD-L1 in h-HER2-MB49 tumors, showing significant differences between treatment groups. (F) Representative flow cytometry profiles detecting CD8 (CTL marker), GZMB, and IFNγ, markers of T cell activity, in h-HER2-MB49 tumors from different treatment groups. (G, H) Quantification of CD8+GZMB+/IFNγ+ CTLs and CD8+/CD3+cell percentages in tumor masses from the different treatment groups (n=5 mice per group). Data are presented as mean±SD, *p<0.05, **p<0.01, ***p<0.001. ADC, antibody-drug conjugate; BCa, bladder cancer. PBS, phosphate buffered saline.

Article Snippet: The primary antibodies included: Anti-PD-L1 antibody ( EPR19759 ), abcam, ab213524; anti-PD-L1 antibody ( EPR20529 ), abcam, ab213480; anti-CD8 alpha antibody ( EPR21769 ), abcam, ab217344; anti-CD8 alpha antibody (CAL66), abcam, ab237709; Granzyme B Polyclonal antibody, Proteintech, 13588-1-AP; TAZ Rabbit Polyclonal Antibody, Proteintech, 23306-1-AP.

Techniques: Injection, Immunofluorescence, Staining, Flow Cytometry, Marker, Activity Assay, Saline

Journal: Cell Genomics

Article Title: Predominant mutated non-canonical tumor-specific antigens identified by proteogenomics demonstrate immunogenicity and tumor suppression in CRC

doi: 10.1016/j.xgen.2025.101062

Figure Lengend Snippet: Non-canonical neo-antigens presented by MHC class I effectively activate CD8 + T cells and suppress tumor growth in a mouse model (A) Workflow for identification and validation of neo-epitopes in a mouse model. (B) Schematic of peptide challenge assay. (C and D) Spot-forming units (SFU; IFN-γ spots per 1 × 10 5 cells) for neo-epitopes derived from intronic (C) and intergenic (D) regions at 14 days post immunization. PBS served as the negative control. (E) Schematic of the in vivo anti-tumor experiment. All ELISpot-confirmed neo-antigens were combined into a multi-epitope vaccine. (F) Tumor volume changes following subcutaneous MC38 cell injection. (G) Tumor-volume changes after antibody-mediated blockade of CD8 + T cells, CD4 + T cells, NK cells, or macrophages. (H) Immunostaining of tumor sections showing CD8 + T, CD4 + T, and regulatory T (Treg) cell infiltration across treatment groups (PBS, control vaccine “CtrlVax and neo-antigen vaccine “Vax”). Scale bar: 40 μm. (I) UMAP projection of single-cell transcriptomic data, annotating tumor-infiltrating lymphocyte (TIL) subpopulations: cytotoxic CD8 + T cells, exhausted CD8 + T cells, exhausted CD4 + T cells, naive CD4 + T cells, proliferating CD4 + T cells, Tregs, NK cells, and B cells. (J) Dot plot of marker gene expression across TIL subsets. Dot size. fraction of cells expressing the gene; dot color. mean normalized expression. (K) Proportion of CD3 + T cells among total tumor-infiltrating immune cells in treatment groups (PBS, CtrlVax, and Vax). (L) Relative distribution of CD4 + T cell subpopulations across treatment groups (PBS, CtrlVax, and Vax) shown as pie charts. Error bars represent mean ± SEM; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Recombinant Anti-CD8 alpha antibody (Rabbit mAb) , Servicebio , Cat# GB15068;RRID: AB_3246431.

Techniques: Biomarker Discovery, Derivative Assay, Negative Control, In Vivo, Enzyme-linked Immunospot, Injection, Immunostaining, Control, Single Cell, Marker, Gene Expression, Expressing

Journal: Nature

Article Title: Sleep modulates hematopoiesis and protects against atherosclerosis

doi: 10.1038/s41586-019-0948-2

Figure Lengend Snippet: a, Image of a sleep fragmentation cage. b, Body weight (n=10 per group). c, Plasma cholesterol at ZT3 (n=5 per group). d, Plasma glucose at ZT3 (n=5 per group). e , Glucose tolerance test (GTT) beginning at ZT3 and ZT12 (n=4 per group). f-h, Apoe −/− mice were placed in sleep fragmentation chambers where the sweep bar operated during the dark period (ZT12–0) when mice are normally awake. Control mice were maintained in SF chambers with a stationary sweep bar. f , Assessment of atherosclerosis and lesion area (n=5 per group). g, Assessment of blood Ly-6C hi monocytes and neutrophils (n=5 per group). h, Assessment of bone marrow LSKs and proliferation (n=5 per group). i, Aortic macrophage proliferation in Apoe −/− and Apoe −/− SF mice after 16 weeks of SF at ZT3 and ZT14 (n=5 Apoe −/− ; n=4 Apoe −/− SF). j , Quantification at ZT3 in Apoe −/− and Apoe −/− SF mice of B cells, CD4 + T cells and CD8 + T cells in blood (n=10 Apoe −/− ; for B and CD4 T cells n=6 Apoe −/− SF for CD8 T cells n=7 Apoe −/− SF), k, spleen (n=10 Apoe −/− ; n=7 Apoe −/− SF) and l, B cells in bone marrow (n=10 Apoe −/− ; n=7 Apoe −/− SF). Data presented as mean ± s.e.m.

Article Snippet: The following monoclonal antibodies were used for flow cytometric analysis: anti-CD45 (BioLegend, clone30-F11, Cat#103147, Lot#B243834), anti-CD45.1 (BioLegend, clone A20, Cat#110708), anti-CD45.2 (BioLegend, clone 104, Cat#109802), anti-CD3 (BioLegend, clone 17A2, Cat#100206), anti-CD90.2 (BioLegend, clone 53–2.1, Cat#105308, Lot#B260050), anti-CD19 (BioLegend, clone 6D5, Cat#115508, Lot#B226581), anti-B220 (BD Biosciences, clone RA3–6B2, Cat#553089, Lot#6012954), anti-NK1.1 BioLegend, clone PK136, Cat#108708), anti-Ly6G (BioLegend, clone 1A8, Cat127614#, Lot#B259670), anti-Ly6C (BioLegend, AL-21, Cat#128006, Lot#B247728), anti-MHCII (BioLegend, clone M5/114.152, Cat#107602, Lot#B217859), anti-F4/80 (Biolegend, clone BM8, Cat#123114, Lot#B237342), anti-CD11b (BioLegend, clone M1/70, Cat#101226, Lot#B238268), anti-CD115 (BioLegend, clone AFS98, Cat#135517, Lot#B265220), anti-Ter119 (BioLegend, clone TER-119, Cat#116208, Lot#B220899), anti-CD34 (eBioscience, clone RAM34, Cat#11–0341-85, Lot#E00265–1634), anti-CD49b (BioLegend, clone DX5, Cat#1089008, Lot#B258302), ant-CD11c (BioLegend, clone N418, Cat#117310, Lot#B206713), anti-IL7Rα (BioLegend, clone SB/199, Cat#121112, Lot#B189668), anti-CD16/32 (BioLegend, clone 93, Cat#101324, Lot#B250025), anti-CD150 (BioLegend, clone TC15–12F12.2, Cat#115922, Lot#B220585), anti-cKit (BioLegend, clone 2B8, Cat#105814, Lot#B252918), anti-CD135 (BioLegend, clone A2F10, Cat#135310, Lot#B234045), anti-CD48 (BioLegend, clone HM48–1, Cat#103426, Lot#B236445), anti-Sca1 (BioLegend, clone D7, Cat#108126, Lot#B234288), anti-CD8 (BD Bioscience, clone 53–6.7, Cat#553035, Lot#2296946), anti-CD4 (BioLegend, clone GK1.5, Cat#100428, Lot#B237336), anti-SiglecF (BD Pharmingen, clone E50–2440, Cat#562680, Lot#7054789), anti-CXCR4 (Invitrogen, clone 2B11, Cat#12–9991-81, Lot#B251481), anti-CXCR2 (BioLegend, clone SA044G4, Cat#149307, Lot#B251481), anti-BrdU (eBioscience, clone BU20A, Cat#17–5071-42, Lot#4319920).

Techniques:

Journal: Nature Communications

Article Title: Somatic mTOR mutation in clonally expanded T lymphocytes associated with chronic graft versus host disease

doi: 10.1038/s41467-020-16115-w

Figure Lengend Snippet: a Gating strategy in screening experiment using the panel of TCR Vβ antibodies. First, CD3 lymphocyte were separated into CD4 + and CD8 + populations using monoclonal antibodies (CD8 + : Pe-Cy7, CD4 + : PerCP). Next, the samples were stained with Vβ specific antibody cocktails included in IO Test Beta Mark TCR beta Repertoire Kit (Beckman-Coulter Immunotech, USA). The β variable chain family was determined based on FITC and PE positivity from CD4 + and CD8 + populations according to the manufacturer’s instruction. Vβ20 clone was detected from total CD4 + T cells (52.9%, middle panel) and total CD8 + T cells (1.74%, right). b Flow cytometry Vβ screening results from the index patient’s peripheral blood sample. T cell clonality with antibodies which target Vβ region of TCR was analysed of CD4 + T cells. The increased distribution suggests that the cells have large T cell clone. c Increased Vβ20 bearing clonotype over time in the index patient’s CD4 + T cells. Source data are provided as a Source data file. d T cell repertoire of FACS-sorted CD4 + Vβ20+ and CD8 + T cells analysed with TCRβ deep sequencing (Adaptive Biotechnologies). The TCRBV30-01 clone was detected in the CD4 + Vβ20+ fraction, but not in the CD8 + fraction. e Multicolor flow cytometry was applied to identify the immune phenotype of HSCT donor and index patient’s memory T cell subtypes. Central memory (CM), naïve, effector memory (EM), and terminal effector memory (TEMRA) cells. f The relative proportion of granzyme B positive (GrB + ) CD4 + T cells and GrB + CD8 + T cells in index patient. Index patient’s PBMCs were stained with anti-CD45, −CD3, −CD4, and −CD8 (surface markers), and then GrB stained after fixation and permeabilization. Stained cells were analyzed using FACSVerse.

Article Snippet: Thereafter, HIER, peroxide and protein block were repeated, followed by application of anti-CD8 (1:500, Clone: C8/144B, Cat#: BSB 5174, Lot#: 5174JDL05, BioSB) primary antibody, HRP-conjugated secondary antibody diluted 1:3 with washing buffer and TSA 555 (PerkinElmer).

Techniques: Staining, Flow Cytometry, Sequencing

Journal: Nature Communications

Article Title: Somatic mTOR mutation in clonally expanded T lymphocytes associated with chronic graft versus host disease

doi: 10.1038/s41467-020-16115-w

Figure Lengend Snippet: Somatic mutations discovered in CD4 + T cells in the index patient, detected from 2013 sample.

Article Snippet: Thereafter, HIER, peroxide and protein block were repeated, followed by application of anti-CD8 (1:500, Clone: C8/144B, Cat#: BSB 5174, Lot#: 5174JDL05, BioSB) primary antibody, HRP-conjugated secondary antibody diluted 1:3 with washing buffer and TSA 555 (PerkinElmer).

Techniques: Mutagenesis

Journal: Nature Communications

Article Title: Somatic mTOR mutation in clonally expanded T lymphocytes associated with chronic graft versus host disease

doi: 10.1038/s41467-020-16115-w

Figure Lengend Snippet: a Locations of mTOR, TLR2 , and NFκB2 somatic mutations. Linearized structure of MTOR, NFκB2 , and TLR2 presenting the location of somatic mutations. mTOR P2229R mutation is located in the kinase domain, NFκB2 P882Q in the C-terminus, and TLR2 W558L between LRR (Leucine-rich repeats) domain and transmembrane (TM) domain. b A heterozygous mTOR mutation (G to C, P2229R ) was detected in CD4 + T cells by Sanger sequencing. c Variant allele frequencies (VAFs) of mTOR P2229R , NFκB2 P882Q , and TLR2 W558L mutations in the index patient’s CD4 + T cells over time as measured with amplicon sequencing. Source data are provided as a Source data file. d VAFs (%) of mTOR P2229R mutation from the index patient’s skin, liver, and eyes biopsy. e Immunofluorescence staining indicated CD3 + CD4 + and CD3 + CD8 + T cell infiltration in the skin. Paraffin embedded skin biopsy from the index patient was sectioned and stained with antibody specific human CD3 (cyan), CD4 (green), and CD8 (red). White arrows indicate infiltrated CD3 + CD4 + or CD3 + CD8 + T cells. Original imaging magnification: 20×, the figure has been further zoomed to 25× for visualization. Scale bar: 50 μm.

Article Snippet: Thereafter, HIER, peroxide and protein block were repeated, followed by application of anti-CD8 (1:500, Clone: C8/144B, Cat#: BSB 5174, Lot#: 5174JDL05, BioSB) primary antibody, HRP-conjugated secondary antibody diluted 1:3 with washing buffer and TSA 555 (PerkinElmer).

Techniques: Mutagenesis, Sequencing, Variant Assay, Amplification, Immunofluorescence, Staining, Imaging

Journal: Nature Communications

Article Title: Somatic mTOR mutation in clonally expanded T lymphocytes associated with chronic graft versus host disease

doi: 10.1038/s41467-020-16115-w

Figure Lengend Snippet: Somatic MTOR and NFκB2 mutations validated by amplicon sequencing in the index patient.

Article Snippet: Thereafter, HIER, peroxide and protein block were repeated, followed by application of anti-CD8 (1:500, Clone: C8/144B, Cat#: BSB 5174, Lot#: 5174JDL05, BioSB) primary antibody, HRP-conjugated secondary antibody diluted 1:3 with washing buffer and TSA 555 (PerkinElmer).

Techniques: Amplification, Sequencing

Journal: Nature Communications

Article Title: Somatic mTOR mutation in clonally expanded T lymphocytes associated with chronic graft versus host disease

doi: 10.1038/s41467-020-16115-w

Figure Lengend Snippet: Real-time cell analysing (RTCA) systems, xCELLigence TM , was applied to monitor real-time killing effect of primary fibroblasts obtained from the index patient and healthy donors. a Index patient’s primary fibroblasts were cultured as monolayers for 24 h to reach full confluence. Once confluent, the effector cells; NK-92 cell (positive control, 8:1), primary CD4 + T cells (4, 8, 16:1) and primary CD8 + T cells (16:1) with different ratios (effector cells:fibroblasts) were added to each well followed by co-culture (arrow indicates the point of adding the effector cells, 0 h). The control (black line, media) shows the impedance of the fibroblasts without any added effectors. The cell impedance was measured every 30 min for 21 h. The measured impedance was expressed as Cell Index with the normalization performed at time of addition of effector cells. b Visualization of monolayers of the primary fibroblast before and 21 h after addition of effector cells (8:1 for all effector cells). c CD4 + T cells and CD8 + T cells from healthy donors were added to healthy donors’ own fibroblasts ( n = 3). The cell impedance was measured for 21 h. Data is representative of three independent individuals. d Index patient’s fibroblasts were seeded with HLA1 and HLA2 antibodies (10 and 20 μg/mL). After CD4 + T cells (CD4 + T cells:fibroblasts = 8:1) were added the cell impedance was measured for 21 h. Dots represent mean values and error bars indicate range ( n = 2 for all conditions, technical duplicates). Source data are provided as a Source data file.

Article Snippet: Thereafter, HIER, peroxide and protein block were repeated, followed by application of anti-CD8 (1:500, Clone: C8/144B, Cat#: BSB 5174, Lot#: 5174JDL05, BioSB) primary antibody, HRP-conjugated secondary antibody diluted 1:3 with washing buffer and TSA 555 (PerkinElmer).

Techniques: Cell Culture, Positive Control, Co-Culture Assay