Journal: Experimental cell research

Article Title: The CNS microenvironment promotes leukemia cell survival by disrupting tumor suppression and cell cycle regulation in pediatric T-cell acute lymphoblastic leukemia.

doi: 10.1016/j.yexcr.2024.114015

Figure Lengend Snippet: Fig. 3. The role of SETD2 in tumor suppression and apoptosis. A) RNA-sequencing analysis of normalized FPKM values (TARGET) showing decreased expression of SETD2 in CNS3 diagnosis patients compared to CNS1 diagnosis samples. (Significance calculated by one-way ANOVA comparing the mean of all groups to the mean of the control group (CNS1), displayed as adjusted p-value. *p < 0.05). B) RNA-sequencing correlation analysis of SETD2 and TP53 (FPKM) showing a negative correlation (r = −0.694, **p=<0.01) and MDM2 showing a positive correlation (r = 0.623, *p=<0.05) in CNS relapsed T-ALL patients (TARGET). (Correlation was calculated using Pearson correlation coefficients, two-tailed with 95% confidence interval). C) String diagram showing protein-protein interactions directly and indirectly experimentally linking SETD2 with TP53, MDM2, BCLX, BCL2, NANOG, SOX2, OCT4, SIRT1 and EP300. D) mRNA expression levels (fold change versus empty vector control) of MDM2, TP53, MYCC, SOX2, NANOG, and OCT4 24 h after SETD2 overexpression by transfection in MOLT16 cells. E) mRNA expression levels (displayed as normalized fold change) of SETD2 overexpression in T-ALL cell lines MOLT16, SUP-T1, CCL-119 has a negative correlation with TP53 (r = −0.709, p = ns), and a positive correlation with SOX2 (r = 0.881, p = ns), NANOG (r = 0.844, p = ns) and OCT4 (r = 0.896, p = ns). (Correlation was calculated using Pearson correlation coefficients, two-tailed with 95% confidence interval). F) Expression levels of BCL2 and BCLX isoforms after SETD2 overexpression in T- ALL cell lines (MOLT16, SUP-T1 and CCL-119) normalized to empty vector control (Significance calculated by multiple two-tailed, unpaired Student’s t-test with Holm-ˇSíd´ak correction for multiple comparisons, displayed as adjusted p-value, *p < 0.05 **p < 0.01).

Article Snippet: DH5α competent cells (Invitrogen) were transformed by heat shock in 42 ◦C water bath using SETD2 Human Tagged ORF Clone (Origene, RG224760) or pCMV6-AC-GFP Mammalian Expression Vector (Origene, PS100010) followed by overnight culture on LB agar plates (SigmaAldrich) containing 100 μg/ml ampicillin (Sigma-Aldrich) at 37 ◦C.

Techniques: RNA Sequencing, Expressing, Biomarker Discovery, Control, Two Tailed Test, Protein-Protein interactions, Plasmid Preparation, Over Expression, Transfection

Journal: Carcinogenesis

Article Title: Downregulation of hedgehog-interacting protein (HHIP) contributes to hexavalent chromium-induced malignant transformation of human bronchial epithelial cells

doi: 10.1093/carcin/bgaa085

Figure Lengend Snippet: Ectopically expressing HHIP reversed the aberrant activation of Hh signaling pathway in Cr(VI)-transformed cells. Cr(VI)-transformed cells (Cr4) were transfected with pCMV6-HHIP and control vector plasmids, and selected for stable transfectants. (A) Total RNA was extracted from cells stably expressing HHIP (Cr4-HHIP) and its control vector (Cr4-EV) cells. HHIP mRNA expression was detected by reverse transcription-quantitative PCR, and normalized to β-actin mRNA expression. Results were presented as relative fold change to the expression level of Cr4-EV cells. Results are mean ± SD (n = 3). *P < 0.05. (B) Expression of HHIP protein was determined by western blot using antibody specifically against Myc-tag and HHIP. β-Actin was served as the loading control. (C) Cells were seeded in eight-chamber culture slides and analyzed for ectopic HHIP expression by immunofluorescent staining using antibody against Myc-tag. (D) Expression of Hh target genes GLI1 and GLI2 were determined by reverse transcription-quantitative PCR and normalized to mRNA expression of β-actin. Results were presented as relative fold change to Cr4-EV cells. Data were mean ± SD (n = 3). *P < 0.05.

Article Snippet: Cr(VI)-transformed cells stably expressing Myc-tagged HHIP were established by transfecting pCMV6-HHIP (Origene, RC206868) using lipofectamine LTX reagent (Thermo Fisher Scientific, 15338030) and selected with 800 μg/ml G418 (Thermo Fisher Scientific, 10131035).

Techniques: Expressing, Activation Assay, Transformation Assay, Transfection, Plasmid Preparation, Stable Transfection, Real-time Polymerase Chain Reaction, Western Blot, Staining

Journal: Oncogenesis

Article Title: Teriflunomide modulates the PD-1/PD-L1 axis and enhances antitumor immunity in colorectal cancer

doi: 10.1038/s41389-026-00607-3

Figure Lengend Snippet: A CRC cell lines were treated with the indicated concentrations of TER for 72 h. Cell viability assessed using the CCK assay is shown. B Human CRC cell lines were co-cultured with hPD-1 Jurkat-T cells and treated with the indicated concentrations of TER for 72 h. C PD-L1 protein expression in CRC cells co-cultured with hPD-1 Jurkat-T cells and treated with TER for 72 h. GAPDH was used as a loading control. The results are shown as the mean ± SEM. * <0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with the respective control.

Article Snippet: Next, 5 μL of 0.5 mg/mL biotinylated hPD-1 (#71109, BPS Bioscience) was added, and the plates were incubated for 2 h at room temperature.

Techniques: Cell Culture, Expressing, Control

Journal: Oncogenesis

Article Title: Teriflunomide modulates the PD-1/PD-L1 axis and enhances antitumor immunity in colorectal cancer

doi: 10.1038/s41389-026-00607-3

Figure Lengend Snippet: A The viability of hPD-1 Jurkat-T cells and hPD-L1 CHO cells following treatment with the indicated concentrations of TER for 24 h. B Luciferase activity measured using a PD-1/PD-L1 blockade bioassay. hPD-1 Jurkat-T cells (effector cells) were co-cultured with hPD-L1-expressing aAPC/CHO-K1 cells (target cells) in the presence of indicated concentrations of TER. The luminescence signal indicates the level of TCR signaling activation. αPD-L1 was used as a positive control. * <0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with the respective control.

Article Snippet: Next, 5 μL of 0.5 mg/mL biotinylated hPD-1 (#71109, BPS Bioscience) was added, and the plates were incubated for 2 h at room temperature.

Techniques: Luciferase, Activity Assay, Bioassay, Cell Culture, Expressing, Activation Assay, Positive Control, Control

Journal: Oncogenesis

Article Title: Teriflunomide modulates the PD-1/PD-L1 axis and enhances antitumor immunity in colorectal cancer

doi: 10.1038/s41389-026-00607-3

Figure Lengend Snippet: A The viability of hPD-L1 MC38 cells following treatment with the indicated concentrations of TER for 72 h. B CD8 + T cells were isolated from tumors of hPD-1 knock-in mice bearing hPD-L1 MC38 tumors. These tumor-infiltrating CD8 + T cells were co-cultured with hPD-L1 MC38 cells as target cells in the presence of TER for 72 h. Cell viability measured using the CCK assay is depicted. C PD-L1 expression in hPD-L1 MC38 cells, as assessed by western blot analysis using protein lysates from co-culture conditions. GAPDH was used as a loading control. D The levels of immune-related factors, including GrB, IL-2, and IFN-γ, measured in the co-culture supernatant by ELISA. * <0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with the respective control.

Article Snippet: Next, 5 μL of 0.5 mg/mL biotinylated hPD-1 (#71109, BPS Bioscience) was added, and the plates were incubated for 2 h at room temperature.

Techniques: Isolation, Knock-In, Cell Culture, Expressing, Western Blot, Co-Culture Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: Oncogenesis

Article Title: Teriflunomide modulates the PD-1/PD-L1 axis and enhances antitumor immunity in colorectal cancer

doi: 10.1038/s41389-026-00607-3

Figure Lengend Snippet: A Body weight of hPD-1 knock-in mice during the treatment period. The mice were treated with vehicle or TER (10 or 30 mpk) for the indicated time. B Spleen weight of mice at the endpoint of the experiment. C Tumor volume was measured over time in hPD-1 knock-in mice bearing hPD-L1 MC38 tumors treated with vehicle or TER (10 or 30 mpk). Representative images of excised tumors from each group are shown. D Tumor weight at the endpoint of the experiment. E Flow cytometry analysis of CD8 + T-cell populations in tumors from each treatment group. F PD-L1 expression in tumors from each group, as assessed by western blot analysis. GAPDH was used as a loading control. G IHC staining of tumor sections for immune-related markers, including CD8 + T cells and GrB. Representative images from each group are shown, and the quantitation of marker-positive cells per field is presented. * <0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with the respective control.

Article Snippet: Next, 5 μL of 0.5 mg/mL biotinylated hPD-1 (#71109, BPS Bioscience) was added, and the plates were incubated for 2 h at room temperature.

Techniques: Knock-In, Flow Cytometry, Expressing, Western Blot, Control, Immunohistochemistry, Quantitation Assay, Marker

Journal: Oncogenesis

Article Title: Teriflunomide modulates the PD-1/PD-L1 axis and enhances antitumor immunity in colorectal cancer

doi: 10.1038/s41389-026-00607-3

Figure Lengend Snippet: A Body weight of hPD-1 knock-in mice during the treatment period. The mice were treated with vehicle or TER (30 mpk) and received either an isotype control or a CD8 depletion antibody. B Spleen weight of mice at the endpoint of the experiment. C Tumor volume was measured in hPD-1 knock-in mice bearing hPD-L1 MC38 tumors over time following treatment with vehicle or TER (30 mpk) with or without CD8 depletion. Representative images of excised tumors from each group are shown. D Tumor weight at the endpoint of the experiment. E Flow cytometry analysis confirming CD8 + T-cell depletion in tumors from each treatment group. The proportion of CD8 + cells among total live cells was quantified. F IHC staining of tumor sections for CD8 + T cells and GrB. Representative images from each treatment group are shown, and the quantitation of marker-positive cells per field is presented. * <0.05, ** p < 0.01, and **** p < 0.0001 compared with the respective control.

Article Snippet: Next, 5 μL of 0.5 mg/mL biotinylated hPD-1 (#71109, BPS Bioscience) was added, and the plates were incubated for 2 h at room temperature.

Techniques: Knock-In, Control, Flow Cytometry, Immunohistochemistry, Quantitation Assay, Marker