Journal: Cell Death & Disease

Article Title: Oncogenic activation of SMYD3-SHCBP1 promotes breast cancer development and is coupled with resistance to immune therapy

doi: 10.1038/s41419-025-07570-8

Figure Lengend Snippet: a The protein levels of Smyd3 and Shcbp1 in normal mouse mammary glands, and tumors initiated with HP5712 cells in 3 months old FVB virgin mice as shown by Western blots. b The protein levels of Smyd3 and Shcbp1 in HP5712 cells expressing sgSmyd3, OE-Smyd3, or OE-Smyd3/sgShcbp1 by Western blots. Representative tumor images ( c ) and tumor weight plots ( d ) at day 32 from FVB virgin mice implanted with HP5712 parental (HP tumors), OE-Smyd3-HP5712, sgSmyd3-HP5712, and OE-Smyd3/sgShcbp1-HP5712 cells at 1X10 6 cells per mammary fat pad (n = 8 mice/group). e The plot of relative spleen weight from the same cohort of mice in c and d . f The protein levels of Smyd3, H3K4me3, and Shcbp1 in tumors initiated with HP5712 parental, OE-Smyd3-HP5712, sgSmyd3-HP5712, and OE-Smyd3/sgShcbp1-HP5712 cells by Western blots. g The protein levels of pMek, pErk, and Kras in tumors initiated with HP5712 parental, OE-Smyd3-HP5712, sgSmyd3-HP5712, and OE-Smyd3/sgShcbp1-HP5712 cells as shown by Western blots. h tSNE visualized immune cells from normal FVB mouse mammary glands (FVB MG, n = 6 mice), tumors implanted with parental HP5712 cells (HP Vector Ctr), HP-OE-Smyd3, HP-OE-Smyd3/sgShcbp1, and HP-sgSmyd3 from FVB mice (n = 6 mice/group) by CyTOF analysis. Quantifications of PMN-MDSCs and M-MDSCs ( i ), CD4+ and CD8 + T cells ( j ) in CD45+ immune cell populations from the same cohorts of mice in h (n = 3 biological independent samples/per group). k , l The WT-T cells were activated with CD3 and CD28 antibody. Co-cultured the activated T cells without or with MDSCs from the mice implanted with HP parental cells, HP cells expressing Smyd3, or sgSmyd3, or OE-Smyd3/sgShcbp1 for 72 hours and examined T cells proliferation status ( k ) and quantifications ( l ) of data in k (n = 3 mice/group). Representative images of CD8+/PD1+ double positive cells in mammary tissues from the mice with the genotypes of HP-Ctr, HP-OE-Smyd3, HP-sgSmyd3, and HP-OE-Smyd3/sgShcbp1 ( m ) and quantifications of CD8+/PD1 double positive cells from mammary tissues ( n ) in these mice (n = 4 mice/group, and at least 20 images for each sample were counted). Scale bar: 20 μm. arrow heads point to PD1/CD8 positive cell. o Expressions of PD1 from the spleen with implantation of parental HP cells, HP cells with OE-Smyd3, HP-sgSmyd3, and HP-OE-Smyd3/sgShcbp1 in FVB mice as determined by qPCR. p Summary of Smyd3-Shcbp1oncogenic signals shape the TIME.

Article Snippet: In Vivo anti-mouse CD8a , BioX Cell , BP0117 , , .

Techniques: Western Blot, Expressing, Plasmid Preparation, Cell Culture

Journal: Cell Death & Disease

Article Title: Oncogenic activation of SMYD3-SHCBP1 promotes breast cancer development and is coupled with resistance to immune therapy

doi: 10.1038/s41419-025-07570-8

Figure Lengend Snippet: Representative tumor images ( a ), and tumor weight plots ( b ) at day 22 from FVB mice implanted with HP5712 cells at 1×10 6 cells per mammary fat pad, and treatment with PBS, PD1, trametinib (Tra), and PD1+Tra (n = 8 mice/group). c The plot of tumor volume in the processes of treatment in ( a , b ). d The plot of relative spleen weight from the same cohort of mice in a , b . Quantifications of PMN-MDSCs and M-MDSCs ( e ), CD4+ and CD8 + T cells ( f ) in CD45+ immune cell populations from the same cohorts of mice in a , b by CyTOF analysis at D12 (n = 3 mice/group). Quantifications of PMN-MDSCs and M-MDSCs ( g ), CD4+ and CD8 + T cells ( h ) in CD45+ immune cell populations from the same cohorts of mice in a , b by CyTOF analysis at D22 (n = 3 mice/group). i , j Tumor images from the HP Ctr mice, HP mice treated with αPD1 and Tra, and HP mice treated with αPD1 and Tra with depletion of T cell using CD8 antibody ( i ). The plot of tumor weight ( j ) in the same cohort of mice in ( i ) (n = 8 mice in each group). The protein levels of Smyd3 and Shcbp1 in tumors initiated with HP5712 cells and treatment with PBS, αPD1, Tra, and αPD1+Tra in FVB mice at D12 ( k ) and D22 ( l ) as shown by Western blots. The protein levels of pMek, pErk, Kras, and Grb2 in tumors initiated with HP5712 cells and treatment with PBS, αPD1, Tra, and αPD1+Tra in FVB mice at D12 ( m ) and D22 ( n ) as shown by Western blots. Expressions of Smyd3 ( o ) and Shcbp1 ( p ) in G600 cells with the treatment of E2, Tra, and E2 together with Tra at 1 hour, 2 hours, 4 hours, and 24 hours as determined by qPCR.

Article Snippet: In Vivo anti-mouse CD8a , BioX Cell , BP0117 , , .

Techniques: Western Blot

Journal: Cell Death & Disease

Article Title: Oncogenic activation of SMYD3-SHCBP1 promotes breast cancer development and is coupled with resistance to immune therapy

doi: 10.1038/s41419-025-07570-8

Figure Lengend Snippet: Antibodies used for ChIP, IHC, IF, WB and in vivo mouse study.

Article Snippet: In Vivo anti-mouse CD8a , BioX Cell , BP0117 , , .

Techniques: Immunohistochemistry-IF, In Vivo

Journal: Journal of experimental & clinical cancer research : CR

Article Title: Regnase-1 downregulation promotes pancreatic cancer through myeloid-derived suppressor cell-mediated evasion of anticancer immunity.

doi: 10.1186/s13046-023-02831-w

Figure Lengend Snippet: Fig. 6 Suppression of CTLs by CD11b+ MDSCs is responsible for the acceleration of tumor progression by Regnase-1 downregulation. A-D Evaluation of phenotypes of orthotopic syngeneic tumors of WT or Regnase-1 KO murine pancreatic cancer cells. Representative macro images of pancreatic tumors (A). Relative mRNA levels of Cd8a, Ifng, Fasl, and Gzmb (B) (N = 6 per group). Representative images of HE (C, left panel) and CD8a immunostaining (C, right panel) and the number of CD8-positive cells (C, right) (N = 6 per group). Dot plots of CD3+CD8+ cells evaluated by flow cytometry (D, left) and the proportion of CD8 + cells among CD45+ cells (D, right) (N = 3 per group). E–H Evaluation of phenotypes of orthotopic syngeneic tumors of WT or Regnase-1-KO murine pancreatic cancer cells with or without depletion of CD8+ cells upon anti-CD8a antibody or IgG treatment. Experimental schematic (E). Dot plots of CD3+ and CD8.+ cells in WT or Regnase-1-KO syngeneic tumors upon anti-CD8a antibody or IgG treatment evaluated by flow cytometry (F). Tumor weights (G) (N = 6 per group). The relative mRNA levels of Cd8a, Ifng, Fasl, and Gzmb (H) (N = 6 per group). Student’s t test was used to evaluate differences between 2 groups. One-way ANOVA with Tukey’s post hoc test was used to compare differences among 4 groups. *P < 0.05, scale bars: 100 μm (insets)

Article Snippet: BE0061, a fully neutralizing mAb recognizing CD8a, and control IgG were obtained from Bioxcell.

Techniques: Immunostaining, Flow Cytometry

Journal: International Journal of Biological Sciences

Article Title: BRD7 Inhibited Immune Escape in Nasopharyngeal Carcinoma via Inhibiting PD-L1 Expression

doi: 10.7150/ijbs.103703

Figure Lengend Snippet: BRD7 downregulated PD-L1 expression and enhanced the cytotoxicity of T lymphocytes against NPC cells. (A) CIBERSORT determined the proportion of immune cell populations in NPC. (B) CD8 + T cell infiltration plays a vital role in NPC. (C) Correlation analysis between the expression of BRD7 and immune cells abundance. (D) Correlation analysis between BRD7 expression level and immune cell subtypes in GSE102349 of NPC. (E) Clonogenic assays of 5-8F and CNE2 cells stably transfected with BRD7 overexpression and knockdown or empty vector plasmids with or without T cells co-culture and PD-L1 antibody incubation. Atezolizumab: the PD-L1 antibody. (F) CCK-8 assay of 5-8F and CNE2 cells stably transfected with BRD7 overexpression and knockdown or empty vector plasmids with or without T cells co-culture and PD-L1 antibody incubation. Absorbance values were detected at 450 nm. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant. (G) Flow cytometry detecting the apoptosis ratio of CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 overexpression or empty vector plasmids with or without PD-L1 antibody incubation. (H) Flow cytometry detecting the apoptosis ratio of CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 knockdown or empty vector plasmids. (I) Flow cytometry detecting the ratio of PD-1 + CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 overexpression or empty vector plasmids with or without PD-L1 antibody incubation. (J) Flow cytometry detecting the ratio of PD-1 + CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 knockdown or empty vector plasmids. (K) ELISA detecting the IFN-γ content in the culture medium of T cells co-cultured with 5-8F cells stably transfected with BRD7 overexpression or empty vector plasmids and CNE2 cells stably transfected with BRD7 knockdown or empty vector plasmids with or without PD-L1 antibody incubation for 24 hours. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.

Article Snippet: T lymphocytes and tumor cells were cocultured at a ratio of 10:1 for 24 h. T cells were stained with an Annexin V-FITC/PI Apoptosis Detection Kit (Biosharp, China), APC Anti-Human CD3 Antibody (Elabscience, China), PerCP Anti-Human CD8a Antibody (Elabscience, China), and FITC Anti-Human CD279/PD-1 Antibody (Elabscience, China) according to the instructions.

Techniques: Expressing, Stable Transfection, Transfection, Over Expression, Knockdown, Plasmid Preparation, Co-Culture Assay, Incubation, CCK-8 Assay, Flow Cytometry, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Biological Sciences

Article Title: BRD7 Inhibited Immune Escape in Nasopharyngeal Carcinoma via Inhibiting PD-L1 Expression

doi: 10.7150/ijbs.103703

Figure Lengend Snippet: BRD7 could inhibit the immune escape of NPC. (A) Design of the experiment in vivo . (B) Images of the appearance of subcutaneous tumors in mice. n = 5 per group. (C) Statistical analysis of subcutaneous tumor weight. n = 5 per group. (D) Tumor growth curve. n = 5 per group. (E) Relative BRD7 and PD-L1 mRNA levels measured by q-PCR in tumor tissues. n = 5 per group. (F) Western blot analysis of BRD7, PD-L1, and PI3K/AKT pathway molecules in tumor tissues. (G) Representative images of immunohistochemical staining for BRD7, PD-L1, PI3K/AKT pathway molecules, and CD8 expression in tumor tissues. The scale bar is 20 μm. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.

Article Snippet: T lymphocytes and tumor cells were cocultured at a ratio of 10:1 for 24 h. T cells were stained with an Annexin V-FITC/PI Apoptosis Detection Kit (Biosharp, China), APC Anti-Human CD3 Antibody (Elabscience, China), PerCP Anti-Human CD8a Antibody (Elabscience, China), and FITC Anti-Human CD279/PD-1 Antibody (Elabscience, China) according to the instructions.

Techniques: In Vivo, Western Blot, Immunohistochemical staining, Staining, Expressing

Journal: Acta Pharmaceutica Sinica. B

Article Title: Arsenic trioxide-based nanoparticles for enhanced chemotherapy by activating pyroptosis

doi: 10.1016/j.apsb.2025.08.003

Figure Lengend Snippet: Immunomodulatory effects of AsMn/Dz@BSA-FA. Analysis of the proportion of (A) CD11c + CD86 + cells, (B) CD3 + CD4 + cells, and (C) CD3 + CD8 + cells in tumor tissues of each group of mice using flow cytometry. (D) Immunofluorescence detection of changes in CD86, CD11c, CD8, and CD4 in tumor tissue after treatment with different formulations, Scale bar: 100 μm. (E) Expression levels of CD86, CD8, and CD4 in tumor tissue after treatment with different formulations. Changes in (F) TNF- α and (G)IL-6 levels in tumor tissue after treatment with different formulations. Data are presented as mean ± SD ( n = 5). ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.

Article Snippet: APC Anti-Mouse CD11c Antibody, PE Anti-Mouse CD86 Antibody, FITC Anti-Mouse CD3 Antibody, APC Anti-Mouse CD4 Antibody, PE Anti-Mouse CD8a Antibody and Purified Anti-Mouse CD16/32 Antibody were from Elabscience (Wuhan, China).

Techniques: Flow Cytometry, Immunofluorescence, Expressing

Journal: CytoJournal

Article Title: The mechanism of prostaglandin E2 upregulation of programmed death ligand 1 expression promoting immune escape in non-small cell lung cancer

doi: 10.25259/Cytojournal_129_2025

Figure Lengend Snippet: PGE2 upregulates PD-L1 expression in NSCLC and promotes immune escape response. (a-c) PD-L1 expression detected after PTGES overexpression (OE-PTGES) and knockdown (sh-PTGES), compared with respective negative controls (OE-NC or sh-NC). (d and e) Cytotoxicity tested by LDH kit assay after PTGES overexpression (OE-PTGES) and knockdown (sh-PTGES), compared with respective negative controls (OE-NC or sh-NC). (f and g) CD8 + T cell viability tested after PTGES overexpression (OE-PTGES) and knockdown (sh-PTGES), compared with respective negative controls (OE-NC or sh-NC). (h and i) CD8 + T cell apoptosis examined after PTGES overexpression (OE-PTGES) and knockdown (sh-PTGES), compared with respective negative controls (OE-NC or sh-NC). (j-m) IFN-γ, TNF-α, granzyme B, and perforin quantification by ELISA after PTGES overexpression (OE-PTGES) and knockdown (sh-PTGES), compared with respective negative controls (OE-NC or sh-NC). n = 6; ✶ P < 0.05, ✶ ✶ P < 0.01, ✶ ✶ ✶ P < 0.001. PEG2: Prostaglandin E2, PD-L1: Programmed death ligand 1, NSCLC: Non-small cell lung cancer, PTGES: Prostaglandin E synthase, OE-NC: Overexpression negative control, sh-NC: Short hairpin negative control, LDH: Lactate dehydrogenase, OE-PTGES: Overexpression prostaglandin E synthase, sh-PTGES: Short hairpin prostaglandin E synthase, IFN-γ: Interferon-gamma, TNF-α: Tumor necrosis factor-alpha, ELISA: Enzyme-linked immunosorbent assay.

Article Snippet: First, a single-cell suspension was prepared and incubated with CD3 (E-AB-F1013E, Elabscience, Wuhan, China) and CD8 (E-AB-F1104Q, Elabscience, Wuhan, China) antibodies in the dark.

Techniques: Expressing, Over Expression, Knockdown, Enzyme-linked Immunosorbent Assay, Negative Control

Journal: CytoJournal

Article Title: The mechanism of prostaglandin E2 upregulation of programmed death ligand 1 expression promoting immune escape in non-small cell lung cancer

doi: 10.25259/Cytojournal_129_2025

Figure Lengend Snippet: PGE2 promotes immune escape in NSCLC in vivo by upregulating PD-L1 expression. (a) Isolated tumor images after PTGES overexpression and knockout. (b and c) Changes in tumor weight and volume after PTGES overexpression and knockout (significant difference markers marked with ✶ represent OE-NC versus OE-PTGES, and those marked with # represent sh-NC vs. sh-PTGES). (d-f) WB analysis of PTGES and PD-L1 after PTGES overexpression and knockout in vivo . (g and h) IHC analysis of CD8 after PTGES overexpression and knockout in vivo (scale bar: 20 μm, magnification, 400×). (i-l) IFN-γ, TNF-α, granzyme B, and perforin quantification by ELISA after PTGES overexpression and knockout in vivo . n = 5; ✶ P < 0.05, ✶ ✶ P < 0.01, ✶ ✶ ✶ P < 0.001, ## P < 0.01. OE-NC: Overexpression negative control, sh-NC: Short hairpin negative control. PEG2: Prostaglandin E2, PD-L1: Programmed death ligand 1, NSCLC: Non-small cell lung cancer, PTGES: Prostaglandin E synthase, OE-NC: Overexpression negative control, sh-NC: Short hairpin negative control, OE-PTGES: Overexpression prostaglandin E synthase, sh-PTGES: Short hairpin prostaglandin E synthase, IHC: Immunohistochemistry, IFN-γ: Interferon-gamma, TNF-α: Tumor necrosis factor-alpha, ELISA: Enzyme-linked immunosorbent assay.

Article Snippet: First, a single-cell suspension was prepared and incubated with CD3 (E-AB-F1013E, Elabscience, Wuhan, China) and CD8 (E-AB-F1104Q, Elabscience, Wuhan, China) antibodies in the dark.

Techniques: In Vivo, Expressing, Isolation, Over Expression, Knock-Out, Enzyme-linked Immunosorbent Assay, Negative Control, Immunohistochemistry

Journal: BJC reports

Article Title: A detoxified TLR4 agonist inhibits tumour growth and lung metastasis of osteosarcoma by promoting CD8+ cytotoxic lymphocyte infiltration.

doi: 10.1038/s44276-024-00120-3

Figure Lengend Snippet: Fig. 3 Lipo-MP-LPS induces immunological changes in both primary tumours and lung metastases. a, c Representative immunohis- tochemistry images of xenograft tumours and lung metastases stained with mouse specific anti-CD8a antibodies. Scale bar = 50 μm. b, d Quantification of CD8a positive cells in xenograft tumours and lung metastases. Data are represented as means ± SD; *p < 0.05, using Mann–Whitney U test. e, g Representative immunohistochemistry images of xenograft tumours and lung metastases stained with mouse specific anti-F4/80 antibodies. Scale bar = 50 μm. f, h Quantification of F4/80 positive cells in xenograft tumours and lung tissues. Data are represented as means ± SD; n = 6 mice per group, *p < 0.05, using Mann–Whitney U test. (i) iNOS, MHC II, TNF-α, CD206, and IL10 gene expression in xenograft tumours and lung metastases determined by RT-qPCR. Data are presented as means ± SD; n = 6 mice per group, *p < 0.05, **p < 0.01, using two-tailed t-test. NS not significant.

Article Snippet: In vivo depletion of CD8+ T cells Mice were depleted of CD8+ T cells by the intraperitoneal administration of an anti-CD8a antibody (YTS169.4, rat IgG2b; BioXCell, West Lebanon, NH, USA).

Techniques: Staining, MANN-WHITNEY, Immunohistochemistry, Gene Expression, Quantitative RT-PCR, Two Tailed Test

Journal: BJC reports

Article Title: A detoxified TLR4 agonist inhibits tumour growth and lung metastasis of osteosarcoma by promoting CD8+ cytotoxic lymphocyte infiltration.

doi: 10.1038/s44276-024-00120-3

Figure Lengend Snippet: Fig. 4 The tumour suppressive effect of Lipo-MP-LPS depends on CD8+ T cells. a Representative images of IHC staining for CD8a positive cells of xenograft tumours from mice treated with anti-CD8a antibody or IgG2b isotype. Scale bar = 100 μm. b CD8a expression in spleen tissues determined by RT-qPCR. Data are represented as means ± SD; n = 4. ***p < 0.001, using two-tailed t-test. c Tumour growth curve in C3H/HeN mice treated with empty liposome or Lipo-MP-LPS in addition to injection with anti-CD8a or isotype IgG2b antibodies (n = 6 mice per group). Data are shown as mean tumour volume ± SD. *p < 0.05, using Mann–Whitney U test. d Kaplan–Meier survival curves. p < 0.05 was considered significant using the log-rank test. Representative images of H&E staining of xenograft tumours (e) and lungs (g) for each group. Scale bar = 200 μm. f Quantification of necrotic areas in tumour tissues. h Quantification of lung metastasis areas. Data are presented as means ± SD; *p < 0.05, **p < 0.01; NS not significant, using Mann–Whitney U test.

Article Snippet: In vivo depletion of CD8+ T cells Mice were depleted of CD8+ T cells by the intraperitoneal administration of an anti-CD8a antibody (YTS169.4, rat IgG2b; BioXCell, West Lebanon, NH, USA).

Techniques: Immunohistochemistry, Expressing, Quantitative RT-PCR, Two Tailed Test, Injection, MANN-WHITNEY, Staining

Journal: BJC reports

Article Title: A detoxified TLR4 agonist inhibits tumour growth and lung metastasis of osteosarcoma by promoting CD8+ cytotoxic lymphocyte infiltration.

doi: 10.1038/s44276-024-00120-3

Figure Lengend Snippet: Fig. 6 Survival analysis of patients with osteosarcoma according to immune cell infiltration levels estimated by consensus TME. Of the 84 patients with osteosarcoma, the top third (28 patients) and bottom third (28 patients) were classified into high and low score groups, respectively. Kaplan–Meier curves of (a) CD8+ T cells (b) macrophage, (c) M1 macrophage, and (d) M2 macrophage infiltration for OS (left) and PFS (right) are presented. p < 0.05 was considered significant using the log-rank test.

Article Snippet: In vivo depletion of CD8+ T cells Mice were depleted of CD8+ T cells by the intraperitoneal administration of an anti-CD8a antibody (YTS169.4, rat IgG2b; BioXCell, West Lebanon, NH, USA).

Techniques: