Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: A IOD of ACAT2 expression in CC tissues and adjacent tissues was examined using immunohistochemical staining ( n = 47 biologically independent samples). IOD of DHCR7 B and MSMO1 C expression in CC patients with high ( n = 27 biologically independent samples) or low ( n = 20 biologically independent samples) expression of ACAT2 was examined using immunohistochemical staining. The number of activated CD8 T cells (CD8A + GZMB + ) D or activated NK cells (CD56 + GZMB + ) E infiltrated in the tumor tissues of patients with high ( n = 27 biologically independent samples) and low ACAT2 ( n = 20 biologically independent samples) expression was detected. Data represent mean ± SEM. Statistical analysis was performed using the paired A or unpaired ( B – E ) t-test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: ACAT2 expression in HCeEpiC and CC cell lines was examined using RT-qPCR A and Western blot analysis B ( n = 5 independent experiments). C ACAT2, DHCR7, and MSMO1 expression in CC cells after infection with Scramble-sh, ACAT2-sh #1, and ACAT2-sh #2 was examined using Western blot analysis ( n = 5 independent experiments). D Detection of total cholesterol, free cholesterol, and cholesteryl ester levels in CC cells ( n = 5 independent experiments). The proliferation of CC cells was examined using CCK8 ( E ) and colony formation assays F (n = 5 independent experiments). G CC cells were co-cultured with (E: T = 3:1) with NK cells or CD8 T cells for 6 h, respectively, and the death of CC cells was detected ( n = 5 independent experiments). H IFN-γ and GZMB released from immune cells in a co-culture system with CC cells were examined using ELISA ( n = 5 independent experiments). Data represent mean ± SEM. Statistical analysis was performed using the one-way ( A , B ) or two-way ( C - H ) ANOVA, followed by Tukey’s multiple comparisons test ( A – H ).

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Infection, Cell Culture, Co-Culture Assay, Enzyme-linked Immunosorbent Assay

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: A ACAT2 knockdown efficiency in U14 cells was examined using western blot analysis ( n = 10 independent experiments). B Volume changes of transplanted tumors in mice subcutaneously inoculated with U14 cells (n = 10 animals). C The images and weight of the tumors harvested on day 21 ( n = 10 animals). D Protein expression of ACAT2, MSMO1, DHCR7, and PCNA in transplanted tumors was examined using western blot analysis ( n = 10 animals). E Detection of total cholesterol, free cholesterol, and cholesteryl ester levels in transplanted tumors ( n = 10 animals). The gating strategy for GZMB + NK cells and CD8 + T cells F and quantification G were analyzed using flow cytometry ( n = 10 animals). H Survival of mice over 60 days after subcutaneous inoculation of U14 cells was analyzed using the log-rank test ( n = 20 animals). Data represent mean ± SEM. Statistical analysis was performed using the one-way ( A , C , E , G ) or two-way ( B , D ) ANOVA, followed by Tukey’s multiple comparisons test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Knockdown, Western Blot, Expressing, Flow Cytometry

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: The proliferation of CC cells was examined using CCK8 A and colony formation assays B ( n = 5 independent experiments). C CC cells were co-cultured with (E: T = 3:1) with NK cells or CD8 + T cells, and the death of CC cells was detected ( n = 5 independent experiments). D IFN-γ and GZMB released from immune cells in a co-culture system with CC cells were examined using ELISA ( n = 5 independent experiments). E TGF-β1 released by CC cells was examined using ELISA ( n = 5 independent experiments). F PD-L1 expression levels in CC cells were observed using immunofluorescence staining ( n = 5 independent experiments). Data represent mean ± SEM. Statistical analysis was performed using the two-way ( A – F ) ANOVA, followed by Tukey’s multiple comparisons test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Cell Culture, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Expressing, Immunofluorescence, Staining

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: A Volume changes of transplanted tumors in mice subcutaneously inoculated with U14 cells ( n = 5 animals). B The images and weight of the tumors harvested on day 21 ( n = 5 animals). The gating strategy for GZMB + NK cells and CD8 + T cells C and quantification D were analyzed using flow cytometry ( n = 5 animals). Data represent mean ± SEM. Statistical analysis was performed using the one-way ( B , D ) or two-way A ANOVA, followed by Tukey’s multiple comparisons test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Flow Cytometry

Journal: Bioactive Materials

Article Title: Self-assembled PROTACs enable protein degradation to reprogram the tumor microenvironment for synergistically enhanced colorectal cancer immunotherapy

doi: 10.1016/j.bioactmat.2024.09.022

Figure Lengend Snippet: NP-PROTACs degraded STAT3 and β-catenin, improving the TIME in vivo . ( A ) Treatment regimen diagrams (n = 6). ( B and D ) After the mice were sacrificed, the tumors were photographed, and the tumor weights were recorded. ( C and E ) Both tumor volume and body weight were monitored every day. ( F ) Immunofluorescence staining of tumor tissue revealed the downregulated target protein in each treatment group (scale bars = 100 μm). ( G – J ) Western blot analyses showed that micelle treatment inhibited β-catenin and/or STAT3 levels in vivo . ( K – N ) The levels of CD103 in DCs, the levels of granzyme B and IFN-γ in T cells, and the polarization of M2-type macrophages were analyzed through flow cytometry (n = 3). ( O ) The statistics of the above immune cells were presented as bars, and CD4 + T cell, CD8 + T cell, Th1 infiltration, and polarization of M2-type macrophages were also determined by flow cytometry and were presented. The data were presented as the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: The antibody for CD8a (10122-80-100) was from Biogems.

Techniques: In Vivo, Immunofluorescence, Staining, Western Blot, Flow Cytometry

Journal: Bioactive Materials

Article Title: Self-assembled PROTACs enable protein degradation to reprogram the tumor microenvironment for synergistically enhanced colorectal cancer immunotherapy

doi: 10.1016/j.bioactmat.2024.09.022

Figure Lengend Snippet: Schematic diagram of the anti-tumor mechanism of dual-target-degrading NP-PROTACs. ( A ) β-catenin-targeting LAVTAT and STAT3-targeting SAVTAT were conjugated with DSPE-PEG 2000 -SH through disulfide bonds, jointly self-assembling into NP-PROTACs. ( B ) Upon intravenous injection and accumulation in CRC models, NP-PROTACs released LAVTAT and SAVTAT intracellularly due to cleaved disulfide bonds. Co-degradation of β-catenin and STAT3 triggered cell cycle arrest and apoptosis on CRC cells and improved immune microenvironment. Briefly, the cytotoxic function of CD8 + T cells was activated; an increase in the ratio of M1-type macrophages and a decrease in the ratio of M2-type macrophages were observed; the infiltration of CD103 + DC cells was increased.

Article Snippet: The antibody for CD8a (10122-80-100) was from Biogems.

Techniques: Injection

Journal: Pharmacological research

Article Title: USP19 deficiency enhances T-cell-mediated antitumor immunity by promoting PD-L1 degradation in colorectal cancer.

doi: 10.1016/j.phrs.2025.107668

Figure Lengend Snippet: Fig. 5. PD-L1 reconstitution reverses USP19 deficiency-induced antitumor immunity. A, B PD-L1 reconstituted in USP19 depletion RKO cells was co-cultured with activated T cells for 48 hours, crystal violet (A) and CCK-8 (B) showed that PD-L1 reconstituted promoted RKO cell viability. C, D Annexin V-FITC and pro- pidium iodide (PI) apoptosis assays showed that PD-L1 reconstituted reversed the apoptosis ratio mediated by USP19 deficiency. E Western blot analysis showed that PD-L1 reconstituted reversed the cleaved caspase-3 protein expression mediated by USP19 deficiency. F-H Flow cytometry revealed that PD-L1 reconstitution reversed the USP19 deficiency-induced increase in the secretion of IFN-γ and GzmB by CD8+ T cells. Data in (B, D, G, H) are shown as mean ± SD. Statistical significance was assessed using one-way and two-way ANOVA (*p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: GAPDH (cat# AC002, ABclonal), USP19 (cat# 25768–1-AP, Proteinch), Myc (cat# M192–3, MBL), PD-L1 (cat# 28076–1-AP, Proteinch), HA (cat# M180–3, MBL), Flag (cat #M185–3L, MBL), Lymphocyte separation medium (cat# 7111012, Dakewe), PE/Cyanine7 anti-human CD8a (cat# 300914, BioLegend), FITC anti-human CD3 (cat# 300306, BioLegend), APC anti-human IFN-γ (cat# 502512, BioLegend), Brilliant Violet 421TM anti-human/mouse Granzyme B Recombinant (cat# 396414, BioLegend), Zombie AquaTM Fixable Viability Kit (cat# 423101,BioLegend), Brefeldin A Solution (1,000X) (cat# 420601, BioLegend), Fixation Buffer (cat# 420801, BioLegend), Intracellular Staining Perm Wash Buffer (10X) (cat# 421002, BioLegend), Rat IgG2b isotype control-InVivo (cat# A2116, selleck), InVivcMAb anti-mouse PD-L1 (cat# 10 F.9G2, BioXCell), FITC Anti-Mouse CD4 Antibody [GK1.5] (cat# E-AB-F1097C, Elabscience), PerCP/Cyanine5.5 Anti-Mouse CD3 Antibody (cat# E-AB-F1013J, Elabscience), PE AntiMouse CD3 Antibody [17A2] (cat# E-AB-F1013D, Elabscience), APC Anti-Mouse CD8a Antibody [53–6.7] (cat# E-AB-F1104E, Elabscience), PE/Cyanine7 Anti-Mouse CD45 Antibody (cat# E-AB-F1136H, Elabscience), Caspase-8 Rabbit mAb (cat# 4790, CST), Cleaved Caspase-8 Rabbit mAb (cat# 9496, CST), Cleaved Caspase-9 Rabbit mAb (cat# 7237, CST), Caspase-9 Rabbit mAb (cat# A119654, ABclonal), Caspase3 Rabbit pAb (cat# A0214, ABclonal), Cleaved Caspase-3 (5A1E) Rabbit mAb (cat# 9664, CST), PARP (46D11) Rabbit mAb (cat# 9532, CST).

Techniques: Cell Culture, CCK-8 Assay, Western Blot, Expressing, Flow Cytometry

Journal: Drug Delivery and Translational Research

Article Title: Self-assembled peptide/polymer hybrid nanoplatform for cancer immunostimulating therapies

doi: 10.1007/s13346-023-01410-y

Figure Lengend Snippet: Scheme of the technological strategy: peptide diblocks consisting of PADRE/MAGE-A3 are self-assembled as nanoparticles. These immunoactive nanoparticles can interact with dendritic cells to activate CD4 + and CD8 + T-cells capable of killing cancer cells

Article Snippet: Anti CD4-APC, anti CD25-PE, anti CD8a-APC, and anti CD28-PE antibodies were obtained from Tonbo Biosciences (California, USA).

Techniques:

Journal: Drug Delivery and Translational Research

Article Title: Self-assembled peptide/polymer hybrid nanoplatform for cancer immunostimulating therapies

doi: 10.1007/s13346-023-01410-y

Figure Lengend Snippet: In vitro immune activation studies. a CD80 + and b CD83 + dendritic cell phenotype by incubation of iDCs with nanoparticles at 10 µM for 48 h. Data are shown as the ratio (%) between the mean fluorescence intensity (MFI) of the corresponding marker in iDCs incubated with the different nanoparticles versus the MFI of iDC incubated in culture media (mean ± S.E., n = 4). Data were analyzed with one-way ANOVA, Sidak test, P < 0.05. The capacity of the iDCs treated by nanoparticles to activate allogeneic CD4 + ( c ) and CD8 + ( d ) T lymphocytes was determined by flow cytometry and quantifying the upregulation of CD25 ( c ) and CD28 ( d ), respectively. Results are shown as the ratio between the total event number of CD4 + CD25 + (CD8 + CD28 + T) cells using iDCs incubated with nanoparticles versus total event number of CD4 + CD25 + (CD8 + CD28 +) T cells using iDCs incubated in culture media (mean ± S.E., n = 4). Data were analyzed with one-way ANOVA, Tukey test, P < 0.05

Article Snippet: Anti CD4-APC, anti CD25-PE, anti CD8a-APC, and anti CD28-PE antibodies were obtained from Tonbo Biosciences (California, USA).

Techniques: In Vitro, Activation Assay, Incubation, Fluorescence, Marker, Flow Cytometry