t98g cell lines  (ATCC)

Journal: Translational Oncology

Article Title: The brain imaging feature-related gene NRP2 drives the malignant progression of glioblastoma through the FAK pathway: a Mendelian randomization study

doi: 10.1016/j.tranon.2026.102732

Figure Lengend Snippet: NRP2 mediates the FAK signaling pathway to regulate the malignant progression of GBM cells. (A): NRP2 expression levels in patients from the GSE4290 and GSE68848 data sets. (B): The knockdown effect of sh-NRP2 in U251 and T98G cells was detected by qRT-PCR. (C): WB was used to detect the expressions of pY297-FAK and FAK. (D): The CCK-8 method was used to detect the cell viability of GBM cells in different treatment groups (sh-NC, sh-NRP2, sh-NRP2 + adhesamine). Adhesamine is a FAK activator. (E): The colony formation experiment was used to detect the number of colony formations of GBM cells in different treatment groups; (F): The scratch healing experiment was used to detect the migration ability of GBM cells in different treatment groups; (G): The Transwell experiment was used to detect the invasion ability of GBM cells in different treatment groups; (H): The Annexin V/PI double staining method was used to detect the apoptosis level of cells in different treatment groups. (I): Western blot was used to detect the protein expression of pro-apoptotic indicators clever-caspase3, clever-PARP, BAX and pro-apoptotic indicator BCL2 in different treatment groups. * indicates P < 0.05, **** indicates P < 0.0001.

Article Snippet: T98G cells were cultured in MEM medium containing 10 % FBS (ATCC, 30–2003, USA).

Techniques: Expressing, Knockdown, Quantitative RT-PCR, CCK-8 Assay, Migration, Double Staining, Western Blot

Journal: Translational Oncology

Article Title: The brain imaging feature-related gene NRP2 drives the malignant progression of glioblastoma through the FAK pathway: a Mendelian randomization study

doi: 10.1016/j.tranon.2026.102732

Figure Lengend Snippet: NRP2 promotes the malignant progression of GBM by activating FAK. U251 and T98G cell groups: sh-NC, sh-NRP2, sh-NRP2 + oe-FAK. (A): qRT-PCR was used to detect the mRNA level of FAK; (B): WB was used to detect the protein expression of pY397-FAK and FAK; (C): CCK-8 was used to detect cell viability; (D): Scratch test was used to detect the migration ability of cells; (E): Transwell test was used to detect the invasion ability of cells; (F): WB was used to detect the protein expression of apoptotic proteins clever-caspase3, clever-PARP, BAX and anti-apoptotic protein BCL2. * indicates P < 0.05.

Article Snippet: T98G cells were cultured in MEM medium containing 10 % FBS (ATCC, 30–2003, USA).

Techniques: Quantitative RT-PCR, Expressing, CCK-8 Assay, Migration

Journal: Translational Oncology

Article Title: The brain imaging feature-related gene NRP2 drives the malignant progression of glioblastoma through the FAK pathway: a Mendelian randomization study

doi: 10.1016/j.tranon.2026.102732

Figure Lengend Snippet: The nude mouse experiment demonstrates that NRP2 mediates the Focal-adhesion pathway to regulate the formation of GBM tumors. (A): Nude mice were subcutaneously injected with sh-NRP2 and sh-NC stably transfected T98G cells. Comparison of tumor size in sh-NRP2 nude mice treated with FAK activator adhesamine. (B): Tumor weights of different treatment groups of nude mice. (C): Tumor volumes of different treatment groups of nude mice. (D): IHC detection of the expression of Ki67, BCL2 and BAX in tumor tissues; (E): WB detection of the expression of pY297-FAK and FAK in tumor tissues.

Article Snippet: T98G cells were cultured in MEM medium containing 10 % FBS (ATCC, 30–2003, USA).

Techniques: Injection, Stable Transfection, Transfection, Comparison, Expressing

Journal: Pharmaceuticals

Article Title: Killing Glioblastoma Cells with Glycosylated Indolocarbazole-Based Derivative LCS1269: A Potential Crosstalk Between Micronuclei Formation and the Concurrent Induction of Apoptosis, Necroptosis, and Pyroptosis

doi: 10.3390/ph19040535

Figure Lengend Snippet: LCS1269 reduces the mitochondrial membrane potential. ( A ) The chemical structure of LCS1269. ( B ) The fluorescence intensity of JC-1 aggregates (red) and JC-1 monomers (green) in U87, U251, and T98G cells that were exposed to LCS1269 (2.5 µM) for 24 h (magnification, 400×). The scale bar = 25 µm. ( C ) LCS1269 (2.5 µM) mediates the MMP depolarization (ΔΨm) in U87 cells in a time-dependent manner. ( D ) The time course of a ΔΨm decrease is shown in the associated bar charts. The data are presented as mean ± SD of at least three separate experiments. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The U87, U251, and T98G GBM cells, as well as the mammary gland adenocarcinoma cell line MCF-7, were purchased from ATCC (Manassas, VA, USA) and cultivated as described previously [ ].

Techniques: Membrane, Fluorescence

Journal: Pharmaceuticals

Article Title: Killing Glioblastoma Cells with Glycosylated Indolocarbazole-Based Derivative LCS1269: A Potential Crosstalk Between Micronuclei Formation and the Concurrent Induction of Apoptosis, Necroptosis, and Pyroptosis

doi: 10.3390/ph19040535

Figure Lengend Snippet: LCS1269 triggers lytic cell death. ( A ) The MTT viability assay of T98G cells that were treated with or without LCS1269 (2.5 µM) for 72 h shows that pre-treatment (for 1 h) with the pan-caspase inhibitor Q-VD-OPh (25 µM) partially prevents LCS1269 cytotoxicity. ( B ) The flow cytometry analysis of the LCS1269-exposed T98G cells that were pre-treated with or without Q-VD-OPh (25 µM) for 1 h, followed by Annexin V-FITC/PI double staining. ( C ) The quantification of necrotic (Q1), late apoptotic (Q2), live (Q3), and early apoptotic (Q4) cell populations. ( D ) LCS1269 induces the release of lactate dehydrogenase (LDH) from U87 ( left ), U251 ( middle ), and T98G ( right ) cells in a dose- and time-dependent manner. ( E ) The Hoechst 33258 plus propidium iodide (PI) staining of U251 and T98G cells that either were or were not exposed to LCS1269 (2.5 µM) for 48 h (magnification, 400×). The scale bar = 25 µm. The data are expressed as mean ± SD of at least three separate experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns—not significant.

Article Snippet: The U87, U251, and T98G GBM cells, as well as the mammary gland adenocarcinoma cell line MCF-7, were purchased from ATCC (Manassas, VA, USA) and cultivated as described previously [ ].

Techniques: MTT Viability Assay, Flow Cytometry, Double Staining, Staining

Journal: Pharmaceuticals

Article Title: Killing Glioblastoma Cells with Glycosylated Indolocarbazole-Based Derivative LCS1269: A Potential Crosstalk Between Micronuclei Formation and the Concurrent Induction of Apoptosis, Necroptosis, and Pyroptosis

doi: 10.3390/ph19040535

Figure Lengend Snippet: LCS1269 induces necroptosis followed by NF-κB activation. ( A ) The representative images ( upper ) show the protein levels of p-NF-κB p65 (S536) and the total NF-κB p65 in the U251 cells that were treated with various concentrations of LCS1269 for 24 h ( left ) or at the indicated time points ( right ). The bar plots demonstrate the results of the relative band density of p-NF-κB p65 (S536) and the total NF-κB p65 that was normalized to β-Actin ( lower ). ( B ) The representative images showing the nuclear–cytoplasmic distribution of the total NF-κB p65 in the U251 cells cultured in the absence or presence of LCS1269 (2.5 µM) for 24 h. DAPI was used to label the cell nuclei (magnification, 1000×). The scale bar = 10 µm. ( C ) The reporter assay of NF-κB transcriptional activity in the U251 and T98G cells treated with or without LCS1269 (2.5 µM) for 24 h. The relative NF-κB-Luc activity was calculated in arbitrary units as the ratio of the detected luciferase activity to β-galactosidase activity (β-Gal). ( D ) The Western blotting analysis of p-MLKL (S358) and the total MLKL in the U251 and T98G cells at the indicated time points ( upper ). The bar plots show the results of the relative band density of p-MLKL (S358) and the total MLKL that was normalized to β-Actin ( lower ). ( E ) The representative immunofluorescence images of p-MLKL (S358) distribution in the U251 cells that either were or were not exposed to LCS1269 (2.5 µM) for 24 h, 48 h, and 72 h (white arrows designate the cell membrane localization of p-MLKL (S358)). DAPI was used to counterstain the cell nuclei (magnification, 1000×). The scale bar = 10 µm. ( F ) The MTT assay of the U251 ( upper ) and T98G ( lower ) cells that were pre-treated with the selective MLKL inhibitor necrosulfonamide (NSA) for 1 h and then exposed to LCS1269 for 72 h. β-Actin was used as a loading control. The data are expressed as mean ± SD of at least three separate experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns—not significant.

Article Snippet: The U87, U251, and T98G GBM cells, as well as the mammary gland adenocarcinoma cell line MCF-7, were purchased from ATCC (Manassas, VA, USA) and cultivated as described previously [ ].

Techniques: Activation Assay, Cell Culture, Reporter Assay, Activity Assay, Luciferase, Western Blot, Immunofluorescence, Membrane, MTT Assay, Control

Journal: Pharmaceuticals

Article Title: Killing Glioblastoma Cells with Glycosylated Indolocarbazole-Based Derivative LCS1269: A Potential Crosstalk Between Micronuclei Formation and the Concurrent Induction of Apoptosis, Necroptosis, and Pyroptosis

doi: 10.3390/ph19040535

Figure Lengend Snippet: LCS1269 differentially regulates ZBP1 , AIM2 , and RIPK1 gene expression in the GBM cells. ( A ) The MTT viability assay of the U251 and T98G cells that were pre-incubated for 1 h with the irreversible pan-caspase inhibitor Q-VD-OPh (25 µM), the selective MLKL inhibitor necrosulfonamide (NSA) (5 µM), or both inhibitors simultaneously, followed by the LCS1269 treatment (2.5 µM) for 72 h. ( B ) The qRT-PCR analysis of the mRNA levels of the three PANoptosome genes, ZBP1 , AIM2 , and RIPK1 , in the U87 and U251 cells that were treated with or without LCS1269 at the indicated concentrations for 24 h. ( C ) The qRT-PCR analysis of the time-dependent effects of LCS1269 on ZBP1 , AIM2 , and RIPK1 mRNA expression in the U251 cells. ( D ) The Western blot analysis of the protein levels of p-RIPK1 (S166) and the total RIPK1 in the U87 and U251 cells that were treated with or without LCS1269 at the indicated concentrations for 24 h (upper). The histograms show the densitometric analysis of the band intensity for specific proteins that were normalized to β-Actin (lower). The data are presented as mean ± SD from at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns—not significant.

Article Snippet: The U87, U251, and T98G GBM cells, as well as the mammary gland adenocarcinoma cell line MCF-7, were purchased from ATCC (Manassas, VA, USA) and cultivated as described previously [ ].

Techniques: Gene Expression, MTT Viability Assay, Incubation, Quantitative RT-PCR, Expressing, Western Blot