Journal: iScience

Article Title: Identification of actionable targets for breast cancer intervention using a diversity outbred mouse model

doi: 10.1016/j.isci.2023.106320

Figure Lengend Snippet: TIEG1 regulates tumor immunity and tumor growth. Tumor growth was monitored using Kaplan-Meier tumor-free survival. p < 0.05 was considered significant. (A) C57BL/6 mice and C57BL/6 HER2 Tg mice with (n = 10) or without TIEG1 (n = 9) were electrovaccinated with pE2TM. (B and C) IFN-γ producing T cells were measured by ELISPOT assay. Data are represented as the average and SD in box and whisker plots. (D) Growth of implanted E0771/E2 tumor in HER2 Tg mice with or without TIEG1. (E) Onset age of spontaneous tumors in NeuT mice with or without TIEG1 (n = 9 and 7, respectively). (F) TIEG1 expression measured by qPCR. Data are represented as pooled averages +/− SEM. (G and H) TIEG1/Klf10 ( top panels ) and Cd3d (bottom panel) expression measured by scRNA sequencing.

Article Snippet: Splenocytes were incubated in ELISPOT plates coated with IFN-γ capture mAb for 48 hrs with recombinant HER2 protein (ecd-Fc fusion; SinoBiological).

Techniques: Enzyme-linked Immunospot, Whisker Assay, Expressing, Sequencing

Journal: Cell Communication and Signaling : CCS

Article Title: Tumorous IRE1α facilitates CD8 + T cells-dependent anti-tumor immunity and improves immunotherapy efficacy in melanoma

doi: 10.1186/s12964-024-01470-8

Figure Lengend Snippet: IRE1α signature score is highly correlated with the feature of tumor-infiltrating lymphocytes in melanoma. A Correlation analysis of 38-hub genes representative of the IRE1α signature with tumor-infiltrating lymphocytes score in TCGA SKCM database. B The mRNA expression of Th1-, cytotoxic mechanisms-, chemokines-, T cell, and MHC class I and II genes based on the groups defined relative to IRE1α activity (High or Low) in TCGA SKCM database. C Correlation analysis of XBP1 with PTPRC , CD8A , GZMB and IFN-γ expression in TCGA SKCM database. D Correlation analysis of DNAJB9 and EDEM1 with PTPRC , CD8A and IFN-γ expression in TCGA SKCM database. E Immunohistochemical staining and correlation analysis of XBP1, CD8α and IFN-γ in a cohort of 31 melanoma tissues. Pt.1, Patient 1; Pt.2, Patient 2. Scale bar = 50 μm. F The overall survival of melanoma patients based on the groups defined relative to IRE1α activity (High or Low) in TCGA SKCM database. G The overall survival of melanoma patients with high TIL score (hot tumor) or low TIL score (cold tumor) based on the groups defined relative to IRE1α activity (High or Low) in TCGA SKCM database. r value was calculated by Spearman correlation. p value was calculated by two tailed Student’s t -test

Article Snippet: Heat-mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) was performed, the sections were blocked with 10% goat serum in PBS for 1 h at room temperature and then stained overnight at 4 °C with rabbit pAb to XBP1s (1:100, 24868-1-AP, proteintech), rabbit pAb to Annexin A1 (1:50, 21990-1-AP, proteintech), rabbit pAb to Calreticulin (1:50, 27298-1-AP, proteintech), rabbit pAb to HMGB1 (1:50, 10829-1-AP, proteintech), rabbit mAb to p-MLKL (Ser345) (1:500, 37,333, CST), mouse mAb to Foxp3 (1:50, 65089-1-Ig, proteintech), CoraLite® Plus 488 Rat mAb to CD4 (1:50, CL488-65141, proteintech), rabbit pAb to Granzyme B (1:50, 13588-1-AP, proteintech), rabbit pAb to IFN-γ (1:50, 105,995-T08, Sinobiological) or rat mAb to CD8α (1:50, 65069-1-Ig, proteintech) antibody.

Techniques: Expressing, Activity Assay, Immunohistochemistry, Staining, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: Tumorous IRE1α facilitates CD8 + T cells-dependent anti-tumor immunity and improves immunotherapy efficacy in melanoma

doi: 10.1186/s12964-024-01470-8

Figure Lengend Snippet: Tumorous IRE1α facilitates anti-tumor immunity in a CD8 + T cells-dependent manner. A Schema of the treatment in C57BL/6 mice bearing B16F10 tumors received HA15 treatment as indicated. Tumor burdens, weights and volumes in each group were calculated and displayed in B and C . D Representative flow cytometry data and summary plots of the frequency of CD3 + CD45 + , CD8 + CD3 + , CD11c + CD45 + , Foxp3 + CD4 + and CD8 + T-cells evaluated for expression of Granzyme B and IFN-γ in tumor from xenografts with indicated treatment. E Immunofluorescence staining of XBP1s or CD8α in B16F10 xenografts with or without the treatment of HA15. Scale bar, 50 μm. F - H Scheme representing the experimental procedure ( F ), tumor burdens, tumor weight ( G ), and tumor volume ( H ) of C57BL/6 mice injected subcutaneously with B16F10 tumors with treatment of HA15 and αCD8 depleting antibodies either alone or in combination. Symbols of one dot indicates one mouse, and the error bars are mean with ± SD ( n = 4). Two-tailed Student’s t-test (* p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant)

Article Snippet: Heat-mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) was performed, the sections were blocked with 10% goat serum in PBS for 1 h at room temperature and then stained overnight at 4 °C with rabbit pAb to XBP1s (1:100, 24868-1-AP, proteintech), rabbit pAb to Annexin A1 (1:50, 21990-1-AP, proteintech), rabbit pAb to Calreticulin (1:50, 27298-1-AP, proteintech), rabbit pAb to HMGB1 (1:50, 10829-1-AP, proteintech), rabbit mAb to p-MLKL (Ser345) (1:500, 37,333, CST), mouse mAb to Foxp3 (1:50, 65089-1-Ig, proteintech), CoraLite® Plus 488 Rat mAb to CD4 (1:50, CL488-65141, proteintech), rabbit pAb to Granzyme B (1:50, 13588-1-AP, proteintech), rabbit pAb to IFN-γ (1:50, 105,995-T08, Sinobiological) or rat mAb to CD8α (1:50, 65069-1-Ig, proteintech) antibody.

Techniques: Flow Cytometry, Expressing, Immunofluorescence, Staining, Injection, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: Tumorous IRE1α facilitates CD8 + T cells-dependent anti-tumor immunity and improves immunotherapy efficacy in melanoma

doi: 10.1186/s12964-024-01470-8

Figure Lengend Snippet: Tumorous IRE1α-induced potentiated activation of CD8 + T cells was mediated by NF-κB. A , E A2058 melanoma cells treated with HA15 (10 µM) for 24 h after pre-treated with or without STF-083010 (10 µM) or BAY 11-7085 (1 µM) for 24 h cocultured with or without activated T cell (1:3) for 24 h were subjected to crystal violet staining. Cytotoxicity was quantified by a spectrometer at OD (570 nm) and normalized ratio of cancer cell survival was shown for each well ( n = 4). B-D , F-H Representative flow cytometry data and summary plots of the frequency of CD8 + T cells evaluated for expression of CD69, Granzyme B and IFN-γ in co-culture system with indicated treatment ( n = 4). Data are representative of four independent experiments and shown as mean ± SD. Two-tailed Student’s t-test (* p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant)

Article Snippet: Heat-mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) was performed, the sections were blocked with 10% goat serum in PBS for 1 h at room temperature and then stained overnight at 4 °C with rabbit pAb to XBP1s (1:100, 24868-1-AP, proteintech), rabbit pAb to Annexin A1 (1:50, 21990-1-AP, proteintech), rabbit pAb to Calreticulin (1:50, 27298-1-AP, proteintech), rabbit pAb to HMGB1 (1:50, 10829-1-AP, proteintech), rabbit mAb to p-MLKL (Ser345) (1:500, 37,333, CST), mouse mAb to Foxp3 (1:50, 65089-1-Ig, proteintech), CoraLite® Plus 488 Rat mAb to CD4 (1:50, CL488-65141, proteintech), rabbit pAb to Granzyme B (1:50, 13588-1-AP, proteintech), rabbit pAb to IFN-γ (1:50, 105,995-T08, Sinobiological) or rat mAb to CD8α (1:50, 65069-1-Ig, proteintech) antibody.

Techniques: Activation Assay, Staining, Flow Cytometry, Expressing, Co-Culture Assay, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: Tumorous IRE1α facilitates CD8 + T cells-dependent anti-tumor immunity and improves immunotherapy efficacy in melanoma

doi: 10.1186/s12964-024-01470-8

Figure Lengend Snippet: ER stress inducer enhances the anti-tumor activity of anti-PD-1 antibody in vivo. A Schema of the treatment in C57BL/6 mice bearing B16F10 tumors received HA15 with or without anti-PD-1 antibody combination treatment as indicated. B Images of isolated tumors from mice that received indicated treatment. Tumor volumes and weights in each group were calculated and displayed in C and D . E - H Representative flow cytometry data and summary plots of the frequency of CD3 + CD45 + , CD8 + CD3 + , F4/80 + CD11b + CD45 + , Foxp3 + CD25 + CD4 + and CD8 + T-cells evaluated for expression of Granzyme B and IFN-γ in tumor from xenografts with indicated treatment. Symbols of one dot indicates one mouse, and the error bars are mean with ± S.D ( n = 4). Two-tailed Student’s t-test (* p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant)

Article Snippet: Heat-mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) was performed, the sections were blocked with 10% goat serum in PBS for 1 h at room temperature and then stained overnight at 4 °C with rabbit pAb to XBP1s (1:100, 24868-1-AP, proteintech), rabbit pAb to Annexin A1 (1:50, 21990-1-AP, proteintech), rabbit pAb to Calreticulin (1:50, 27298-1-AP, proteintech), rabbit pAb to HMGB1 (1:50, 10829-1-AP, proteintech), rabbit mAb to p-MLKL (Ser345) (1:500, 37,333, CST), mouse mAb to Foxp3 (1:50, 65089-1-Ig, proteintech), CoraLite® Plus 488 Rat mAb to CD4 (1:50, CL488-65141, proteintech), rabbit pAb to Granzyme B (1:50, 13588-1-AP, proteintech), rabbit pAb to IFN-γ (1:50, 105,995-T08, Sinobiological) or rat mAb to CD8α (1:50, 65069-1-Ig, proteintech) antibody.

Techniques: Activity Assay, In Vivo, Isolation, Flow Cytometry, Expressing, Two Tailed Test

Journal: Vaccines

Article Title: Immunogenicity of the Xcl1 -SARS-CoV-2 Spike Fusion DNA Vaccine for COVID-19

doi: 10.3390/vaccines10030407

Figure Lengend Snippet: Xcl1-S vaccination elicited a significant increase in cellular immune responses. ( A , B ) The number of IFN-γ-expressing cells ( A ) and IL-4-expressing cells ( B ) from splenocytes were measured by ELISpot. Data are mean ± SEM of the spleens from n = 5 mice per group. ( C , D ) Splenocytes were stimulated with a combined mixture of S peptide pools for 18 h. The frequency of S-specific IFN-γ CD4 + and CD8 + T cells was determined using a flow cytometry analysis. Summary graphs showing the frequencies of IFN-γ CD4 + and CD8 + T cells after vaccination. Data shown in the graphs represent the average of five mice in each group, and error bars represent SEM. ★, p < 0.05, ★★, p < 0.01.

Article Snippet: Splenocytes were counted and 2 × 10 5 splenocytes per well were plated into anti-IFN-γ or anti-IL-4 plates and restimulated with the N terminal domain (NTD), receptor binding domain (RBD) and S1 peptides pools with 5 µg/mL concentration for 18 h at 37 °C 5% CO 2 (Sino Biological, cat. PP001, PP002 and PP003-A, Beijing, China).

Techniques: Expressing, Enzyme-linked Immunospot, Flow Cytometry

Journal: Journal of Alzheimer's Disease

Article Title: Evaluation of a DNA Aβ 42 Vaccine in Aged NZW Rabbits: Antibody Kinetics and Immune Profile after Intradermal Immunization with Full-Length DNA Aβ 42 Trimer

doi: 10.3233/JAD-160947

Figure Lengend Snippet: ELISPOT analyzes for Aβ 42 peptide re-stimulated splenocyte cultures of DNA Aβ 42 immunized rabbits. A) The number of IL-17 secreting cells is shown following 48 h in cell culture. B) The number of IFN γ secreting cells. C) The number of IL-4 secreting cells per 10 6 splenocytes. * p < 0.05, ** p < 0.005, **** p < 0.0001.

Article Snippet: ELISPOT assays to determine frequencies of cytokine secreting cells were performed according to standard procedures (48 h), and as previously described using commercial available antibody sets for rabbit IFN γ , IL-17, and IL-4 (Kingfisher Biotech, Saint Paul, MN) [ ].

Techniques: Enzyme-linked Immunospot, Cell Culture

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: (a) Schematic of laser-cut PDMS microwell array reversibly assembled on a polystyrene petri dish and workflow of enzymatically amplified silver metallization using Biotin-BSA and poly-HRP-SA. (b) Cell-phone image of silver metallized wells after assay. (c) Biotin-BSA dilution curve obtained from cell-phone image silver darkness quantification in ImageJ. (d) Assay stack for initial biotinyl tyramide signal amplification testing using biotinylated anti-IFN-γ as capture. (e) Dilution curve of silver darkness of increasing biotinylated anti-IFN-γ as target and 10 µg/mL, 1 µg/mL, and 0 µg/mL biotinyl tyramide. All assays were repeated three (n=3) times and fit lines are four-parameter logistic sigmoidal curves.

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques: Amplification

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: (a) Schematic of immunoassay for cytokine detection. (b) Initial LOD optimization for the IFN-γ assay. (c-e) IFN-γ (n=2), IL-2 (n=5), TNF-α (n=2) dilution curves using recombinant cytokines (f-h) Correlation plots of ELISA absorbance values at 450 nm versus silver darkness values for IFN-γ (f), TNF-α (g), and IL-2 (h). All fit lines are four-parameter logistic sigmoidal curves.

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: (a) Schematic demonstrating the stimulation groups and controls. Dilution curves for IFN-γ (b-c), IL-2 (d-e), and TNF-α (f-g) in pooled LTB+ and LTB-clinical supernatant samples. Clinical samples consisted of NIL (negative control), Ag1, Ag2, and Mitogen (positive control) groups (n=2).

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques: Negative Control, Positive Control

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: Individual participant silver darkness values for IFN-γ (a), IL-2 (b), and TNF-α (c) detection from NIL, Ag1, Ag2, and Mitogen samples of IGRA+ and IGRA-patients using a sample dilution of 1:10. Average of n=2 repeats was plotted for each sample.

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques:

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: (a) Schematic of laser-cut PDMS microwell array reversibly assembled on a polystyrene petri dish and workflow of enzymatically amplified silver metallization using Biotin-BSA and poly-HRP-SA. (b) Cell-phone image of silver metallized wells after assay. (c) Biotin-BSA dilution curve obtained from cell-phone image silver darkness quantification in ImageJ. (d) Assay stack for initial biotinyl tyramide signal amplification testing using biotinylated anti-IFN-γ as capture. (e) Dilution curve of silver darkness of increasing biotinylated anti-IFN-γ as target and 10 µg/mL, 1 µg/mL, and 0 µg/mL biotinyl tyramide. All assays were repeated three (n=3) times and fit lines are four-parameter logistic sigmoidal curves.

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques: Amplification

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: (a) Schematic of immunoassay for cytokine detection. (b) Initial LOD optimization for the IFN-γ assay. (c-e) IFN-γ (n=2), IL-2 (n=5), TNF-α (n=2) dilution curves using recombinant cytokines (f-h) Correlation plots of ELISA absorbance values at 450 nm versus silver darkness values for IFN-γ (f), TNF-α (g), and IL-2 (h). All fit lines are four-parameter logistic sigmoidal curves.

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: (a) Schematic demonstrating the stimulation groups and controls. Dilution curves for IFN-γ (b-c), IL-2 (d-e), and TNF-α (f-g) in pooled LTB+ and LTB-clinical supernatant samples. Clinical samples consisted of NIL (negative control), Ag1, Ag2, and Mitogen (positive control) groups (n=2).

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques: Negative Control, Positive Control

Journal: bioRxiv

Article Title: Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic Metallization

doi: 10.64898/2026.01.27.700981

Figure Lengend Snippet: Individual participant silver darkness values for IFN-γ (a), IL-2 (b), and TNF-α (c) detection from NIL, Ag1, Ag2, and Mitogen samples of IGRA+ and IGRA-patients using a sample dilution of 1:10. Average of n=2 repeats was plotted for each sample.

Article Snippet: Recombinant human IFN-γ protein (11725-HNAS), biotinylated anti-human IFN-γ antibody (11725-R209-B), anti-human IFN-γ antibody (11725-R238), HRP-conjugated anti-human IFN-γ antibody (11725-R209-H), recombinant human TNF-α protein (10602-HNAE), biotinylated anti-human TNF-α antibody (10602-MM08-B), and anti-human TNF-α antibody (10602-MM01) were obtained from SinoBiological.

Techniques: