mcf  (ATCC)

Journal: Genes & Diseases

Article Title: Blockade of co-inhibitory receptor immune checkpoint protein TIM3/CD366 augments the anti-cancer activity of CAR-T therapy in solid tumors: An ovarian cancer example

doi: 10.1016/j.gendis.2025.101978

Figure Lengend Snippet: Specific IFN-γ and TNF-α release of T lymphocytes transduced with TIM-3-silenced HER2-specific chimeric antigen receptor (CAR) or HER2-specific CAR. (A, B) TIM-3-silenced CAR-T cells and control T cells were co-incubated with Galectin-9 + or Galectin-9 – SKOV3 tumor cells (E:T ratio 5:1 or 10:1). At 20 h after coculture, a specific enzyme-linked immunosorbent assay was used to analyze the supernatant for IFN-γ cytokine-release. Results were presented as mean ± standard deviation. (C, D) The detection of TNF-α in the same culture supernatant. Results were presented as mean ± standard deviation. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: Human cervical cancer cell line HeLa, lentivirus packaging cell line HEK 293TD, and human ovarian cancer cell line SKOV3 were purchased from American Type Culture Collection (Manassas, Virginia, USA) and cultured in Dulbecco's modified Eagle's medium (Invitrogen, Grand Island, New York) supplemented with 10% heat-inactivated fetal bovine serum.

Techniques: Transduction, Control, Incubation, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Genes & Diseases

Article Title: Blockade of co-inhibitory receptor immune checkpoint protein TIM3/CD366 augments the anti-cancer activity of CAR-T therapy in solid tumors: An ovarian cancer example

doi: 10.1016/j.gendis.2025.101978

Figure Lengend Snippet: TIM-3 silencing augmented the anti-tumor activity of chimeric antigen receptor-T (CAR-T) cells in vivo . 2 × 10 6 SKOV3 tumor cells expressing luciferase were intraperitoneally inoculated in a xenograft mouse model, and 7 days after inoculation, the 2 × 10 6 HER2-specific CAR-T kdTim-3 cells or CAR-T cells, or untreated T cells were intraperitoneally administered. (A, B) Tumor growth was monitored using an in vivo imaging system. (C) Survival curve of 80-day post-treatment. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: Human cervical cancer cell line HeLa, lentivirus packaging cell line HEK 293TD, and human ovarian cancer cell line SKOV3 were purchased from American Type Culture Collection (Manassas, Virginia, USA) and cultured in Dulbecco's modified Eagle's medium (Invitrogen, Grand Island, New York) supplemented with 10% heat-inactivated fetal bovine serum.

Techniques: Activity Assay, In Vivo, Expressing, Luciferase, In Vivo Imaging

Journal: Poultry Science

Article Title: Houttuynia cordata extract protects against Salmonella infection by targeting type III secretion system 1

doi: 10.1016/j.psj.2026.106937

Figure Lengend Snippet: HCE suppresses the adhesion and invasion of Salmonella in Caco-2 cells. A CCK-8 assay (A) was conducted to evaluate the cytotoxicity of HCE in Caco-2 cells. The effects of HCE on the adhesion (B, D) and invasion (C, E) of ST (B-C) and SP (D-E) to Caco-2 cells were examined at an MOI of 100 (n = 5). Immunofluorescence microscopy was performed to visualize the inhibitory effect of HCE (150 µg/mL) on the invasion of ST (F) and SP (G) in Caco-2 cells. Caco-2 cells infected with the ST Δ invA strain, which was used as a negative control because of its T3SS-1 deficiency showed significantly reduced invasion. ST, Salmonella Typhimurium. SP, Salmonella Pullorum . HCE, Houttuynia cordata extract.

Article Snippet: Caco-2 cells (American Type Culture Collection, ATCC, USA) were cultured in DMEM and seeded into culture plates.

Techniques: CCK-8 Assay, Immunofluorescence, Microscopy, Infection, Negative Control

Journal: Bioactive Materials

Article Title: Harnessing the gut–immune–joint axis: Oral microalgae-based thermoresponsive microspheres enhance intra-articular therapy for rheumatoid arthritis

doi: 10.1016/j.bioactmat.2026.01.037

Figure Lengend Snippet: Evaluation of the protective effects of CG@GelMA on FSN-induced intestinal epithelial barrier disruption in vitro . (A) Immunofluorescence staining of Claudin-1 in Caco-2 monolayers under different treatments. Scale bar: 100 μm. (B) Relative fluorescence intensity of Claudin-1. (C) Immunofluorescence staining of Occludin in Caco-2 monolayers. Scale bar: 100 μm. (D) Relative fluorescence intensity of Occludin. (E) Immunofluorescence staining of ZO-1 in Caco-2 monolayers. Scale bar: 100 μm. (F) Relative fluorescence intensity of ZO-1. (G) Schematic diagram of the Caco-2/RAW 264.7 Transwell co-culture system. (H) Relative TEER of Caco-2 cell monolayers after different treatments. (I) Relative fluorescence intensity of FD4 across Caco-2 monolayers under different treatments. Data are presented as means ± SD. Statistical significance: ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: RAW 264.7 macrophages (Procell, Wuhan, China), Caco-2 intestinal epithelial cells (Pricella, Wuhan, China), and IEC-6 small intestinal epithelial cells (ATCC, USA) were maintained in high-glucose DMEM supplemented with 10 % fetal bovine serum (AiTing, Hangzhou, China) and 1 % penicillin-streptomycin (Gibco, USA), with the medium for IEC-6 cells additionally containing 0.1 U/mL human insulin.

Techniques: Disruption, In Vitro, Immunofluorescence, Staining, Fluorescence, Co-Culture Assay

Journal: STAR Protocols

Article Title: Protocol for enhancing CRISPR-Cas9 genome editing using histone deacetylase inhibition and engineered virus-like particle delivery

doi: 10.1016/j.xpro.2026.104493

Figure Lengend Snippet: FAME-CRISPR improves CRISPR/Cas9 gene editing efficiency through HDACi mediated chromatin relaxation and precise eVLP delivery (A) shows a brief schematic of the major steps required to improve editing efficacy using FAME-CRISPR workflow. (B), (C), (D), and (E) show representative results obtained from MDA-MB-231 one of the utilized cell lines from the original FAME-CRISPR study ([60nM panobinostat] and [1.5 μM belinostat]) (n = 3 biological replicates for all conditions, and error bars reflect standard deviation). Results and raw data for MDA-MB-231 and other tested cell lines are available in the original study and data availability statement section of STAR Protocols.

Article Snippet: MDA-MB-231 , ATCC , #HTB-26.

Techniques: CRISPR, Standard Deviation

Journal: Molecular Therapy Oncology

Article Title: Mitochondrial inhibition enhances the sensitivity of pancreatic ductal adenocarcinoma cells to oncolytic adenovirus

doi: 10.1016/j.omton.2026.201180

Figure Lengend Snippet: Glycolysis inhibitors diminish the virus sensitivity of glycolytic PDAC cells MIA PaCa-2 and PK-59 cells were treated with SCH772984 (SCH) (200 nM) or 2DG (2 mM), followed by infection with OBP-401 (100 MOI) or OBP-702 (10 MOI). (A) Lactate secretion by MIA PaCa-2 and PK-45H cells treated with SCH772984 or 2DG, presented as fold-increase compared with the control group, which was set as 1.0. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (B) Cell lysates of MIA PaCa-2 and PK-45H cells treated with SCH or 2DG for 48 h were subjected to western blot analysis for ERK1/2, GLUT1, and LDHA. (C) MIA PaCa-2 and PK-45H cells were treated with SCH or 2DG, followed by infection with OBP-401 (100 MOI) for 24 or 48 h. Upper panels show representative photographs of immunocytochemical staining for GFP in each group 48 h after infection. Scale bars, 500 μm. Lower graphs show the fluorescence intensity of GFP analyzed under fluorescence microscopy. Data are expressed as mean (SD) of independent experiment ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (D) MIA PaCa-2 and PK-45H cells were co-treated with OBP-702 and SCH772984 or 2DG at the indicated dose for 72 h. Cell viability was quantified using the XTT assay and calculated relative to the mock-infected group. Data are expressed as mean (SD) of independent experiment ( n = 5). The statistical significance of differences between two groups was determined using the Student’s t test. (E) Cell lysates of MIA PaCa-2 and PK-45H cells co-treated with SCH or 2DG and OBP-702 (10 MOI) for 48 h were subjected to western blot analysis for E1A, p53, PARP, and cleaved C-PARP. β-actin was assayed as a loading control. The expression level of each protein was calculated relative to that of mock-treated cells, which was set at 1.0. N.S., not significant; ∗, p < 0.05.

Article Snippet: Two human PDAC cell lines (MIA PaCa-2 and Capan-2) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Virus, Infection, Control, Western Blot, Staining, Fluorescence, Microscopy, XTT Assay, Expressing

Journal: Molecular Therapy Oncology

Article Title: Mitochondrial inhibition enhances the sensitivity of pancreatic ductal adenocarcinoma cells to oncolytic adenovirus

doi: 10.1016/j.omton.2026.201180

Figure Lengend Snippet: p53 activation modulates glutamine metabolism in PDAC cells (A) Glutamine consumption in PDAC cells, presented as fold-increase compared with PBS, which was set as 1.0. (B) Outline of glutamine metabolism, shown from glutamine uptake to α-KG production. (C) Lysates of PDAC cells were subjected to western blot analysis for GDH1/2, OGDH, and IDH1. (D) PDAC cells were infected with OBP-301 or OBP-702 at an MOI of 100 for 48 h. The amount of intracellular α-KG in PDAC cells is shown as fold-increase compared with the mock-infected group, which was set as 1.0. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (E) PDAC cells were infected with OBP-301 or OBP-702 at the indicated MOIs for 72 h. Cell lysates were subjected to western blot analysis for GDH1/2, OGDH, and IDH1. (F) MIA PaCa-2 and PK-59 cells were infected with DL312 or Adp53 at the indicated MOIs for 24 h. The amount of intracellular αKG in PDAC cells is presented as fold-increase compared with mock-infected control groups. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences among four groups was determined using one-way ANOVA followed by Turkey’s multiple comparison procedure. (G) MIA PaCa-2 and PK-59 cells were infected with DL312 or Adp53 at the indicated MOIs for 48 h. Cell lysates were subjected to western blot analysis for p53, GDH1/2, OGDH, and IDH1. β-Actin was assayed as a loading control. The expression level of each protein was calculated relative to that of MIAPaCa-2 cells or mock-treated cells, which was set at 1.0. ∗, p < 0.05.

Article Snippet: Two human PDAC cell lines (MIA PaCa-2 and Capan-2) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Activation Assay, Western Blot, Infection, Control, Comparison, Expressing

Journal: Molecular Therapy Oncology

Article Title: Mitochondrial inhibition enhances the sensitivity of pancreatic ductal adenocarcinoma cells to oncolytic adenovirus

doi: 10.1016/j.omton.2026.201180

Figure Lengend Snippet: Comparison of metabolic phenotypes and virus sensitivity in subcutaneous tumor models with glycolytic and non-glycolytic PDAC cells (A) Representative photographs of immunohistochemical staining for LDHA, GLUT1, and IDH1 in each group. Scale bars, 100 μm. (B) Expression levels of LDHA, GLUT1, and IDH1, calculated by dividing the DAB intensity by the number of cells in randomly selected fields. Data are expressed as mean (SD) of independent experiments ( n = 5). The statistical significance of differences between two groups was determined using the Student’s t test. (C) MIA PaCa-2 tumor-bearing mice received intratumoral injections of PBS (black arrows) or OBP-702 (green arrows) every other day for 3 cycles. Data are expressed as mean (SD) of independent experiments ( n = 5). The statistical significance of differences between two groups was determined using the Student’s t test. (D) PK-59 tumor-bearing mice received intratumoral injections of PBS (black arrows) or OBP-702 (orange arrows). The upper right photographs show tumor-bearing mice in the control and OBP-702-treated groups. The lower right photographs show tumors in the mock and OBP-702 groups. Data are expressed as mean (SD) of independent experiments ( n = 5). The statistical significance of differences between two groups was determined using the Student’s t test. ∗, p < 0.05.

Article Snippet: Two human PDAC cell lines (MIA PaCa-2 and Capan-2) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Comparison, Virus, Immunohistochemical staining, Staining, Expressing, Control

Journal: Molecular Therapy Oncology

Article Title: Mitochondrial inhibition enhances the sensitivity of pancreatic ductal adenocarcinoma cells to oncolytic adenovirus

doi: 10.1016/j.omton.2026.201180

Figure Lengend Snippet: Investigation of the relationship between PET/CT metabolic parameters and glycolytic activity of PDAC tumors (A and B) PET/CT images of MIA PaCa-2 tumor (A) and PK-59 tumor (B). The upper left (a) shows the horizontal section, whereas the lower left (b) shows the sagittal section, and the right (c) shows the coronal section. Dotted circles indicate the tumor area. (C) Comparison of SUVmax values for MIA PaCa-2 and PK-59 tumors. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (D and E) Comparison of MTV (D) and TLG (E) values for MIA PaCa-2 and PK-59 tumors at the indicated thresholds. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (F and G) Scatter diagrams demonstrating correlations between expression of LDHA (F) or GLUT1 (G) and preoperative SUVmax (left), MTV (40%) (center), and TLG (40%) (right) values in patients with PDAC ( n = 30). The statistical significance of the correlations in the scatterplots was determined using Pearson’s correlation analysis. N.S., not significant; ∗, p < 0.05.

Article Snippet: Two human PDAC cell lines (MIA PaCa-2 and Capan-2) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Positron Emission Tomography-Computed Tomography, Activity Assay, Comparison, Expressing