human gbm cell line t98g  (ATCC)

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

Journal: The Journal of Biological Chemistry

Article Title: The role of DNAJC3 in enhancing glioma progression and regulating the tumor immune microenvironment

doi: 10.1016/j.jbc.2025.111059

Figure Lengend Snippet: The ERS-related protein DNAJC3 is significantly overexpressed in glioma. A , volcano map showing the differentially expressed proteins (|log 2 FC| ≥1, and p < 0.01). B , Venn diagram analysis of differentially expressed proteins and ERS-related proteins (relevance score >2). C , expression levels of DNAJC3 in glioma tissues (n = 3, Student's t test, compared with adjacent). D , changes in the expression levels of GRP78 and DNAJC3 in T98G and U251 cells after treatment with different concentrations of tunicamycin (n = 3, ANOVA, compared with 0 μM). The data are presented as mean ± SD. (∗∗∗∗ p < 0.0001; ∗∗∗ p < 0.001; ∗∗ p < 0.01; ∗ p < 0.05). DNAJC3, DnaJ heat shock protein family member C3; ERS, endoplasmic reticulum stress; FC, fold change.

Article Snippet: The human GBM cell lines T98G and U251 were purchased from Procell Life Science & Technology Co, Ltd.

Techniques: Expressing

Journal: The Journal of Biological Chemistry

Article Title: The role of DNAJC3 in enhancing glioma progression and regulating the tumor immune microenvironment

doi: 10.1016/j.jbc.2025.111059

Figure Lengend Snippet: In vitro , overexpression of DNAJC3 promotes the proliferation, migration, invasion, and antiapoptosis ability of T98G and U251 cells. A , effects of high/low expression of DNAJC3 on cell viability were assessed using the CCK-8 assay (n = 3, Student's t test, compared with si-NC/vector). B , the impact of high/low expression of DNAJC3 on cell proliferation capacity was examined through colony formation assay (n = 3, Student's t test, compared with si-NC/vector). C , the influence of high/low expression of DNAJC3 on cell migration ability was evaluated using a wound healing assay (n = 3, ANOVA, compared with si-NC/vector). D , the effect of high/low expression of DNAJC3 on cell migration was assessed using a transwell assay (magnification: 40×) (n = 3, ANOVA, compared with si-NC/vector). E , the impact of high/low expression of DNAJC3 on cell invasion ability was determined using a transwell invasion assay (n = 3, ANOVA, compared with si-NC/vector). F , the effect of high/low expression of DNAJC3 on cell apoptosis was analyzed using flow cytometry (n = 3, Student's t test, compared with si-NC/vector). The data are presented as mean ± SD. (∗∗∗∗ p < 0.0001; ∗∗∗ p < 0.001; ∗∗ p < 0.01; ∗ p < 0.05; CON, control group; DNAJC3, DNAJC3 overexpression group; si-DNAJC3, DNAJC3 knockdown group; si-NC, negative control of the knockdown group; vector, empty vector control of the overexpression group.) CCK-8, Cell Counting Kit-8; DNAJC3, DnaJ heat shock protein family member C3.

Article Snippet: The human GBM cell lines T98G and U251 were purchased from Procell Life Science & Technology Co, Ltd.

Techniques: In Vitro, Over Expression, Migration, Expressing, CCK-8 Assay, Plasmid Preparation, Colony Assay, Wound Healing Assay, Transwell Assay, Transwell Invasion Assay, Flow Cytometry, Control, Knockdown, Negative Control, Cell Counting

Journal: Biomacromolecules

Article Title: Hydrolytically Stable Cationic Bis-MPA Dendrimers as Efficient Transfectants for Glioblastoma Cells and Primary Astrocytes

doi: 10.1021/acs.biomac.5c01202

Figure Lengend Snippet: Direct toxicity of the dendrimers. T98G cells were exposed to increasing dendrimer concentrations ranging from 0.1 to 10 μM and toxicity determined as LDH release to the culture medium determined at 72 h. Data are reported as mean values, with error bars indicating the SEM for n = 8 to 24.

Article Snippet: T98G human GBM cells were obtained from the American Type Culture Collection (ATTC, Rockville, MD, USA) and cultured according to the provider instructions.

Techniques:

Journal: Biomacromolecules

Article Title: Hydrolytically Stable Cationic Bis-MPA Dendrimers as Efficient Transfectants for Glioblastoma Cells and Primary Astrocytes

doi: 10.1021/acs.biomac.5c01202

Figure Lengend Snippet: Cellular uptake of dendrimer/siRNA dendriplexes in T98G cells. (A) T98G cells were incubated for 6 h with dendriplexes formed by FAM-labeled siRNA (100 nM) and dendrimers at the following concentrations: G2-DA (5 μM), G3-DA (0.5 μM), G1-CYS (5 μM), G2-CYS (0.5 μM), or G3-CYS (0.5 μM). Shown are siRNA (FAM), nuclei (Hoechst), brightfield, and merged images. The experiment was performed twice with similar results. (B) Cells were incubated for 6 h with Cy7.5-labeled G3-CYS (1 μM) and Hoechst. Brightfield and merged images are also shown. The experiment was performed twice, with eight frames recorded each time, yielding similar results. (C) Cells were incubated for 6 h with Cy7.5-labeled G3-CYS (1 μM) and confocal images recorded. The figure shows a confocal image of a T98G cell at 2.4 μm above the basal plane, showing intracellular Cy7.5-labeled G3-CYS dendrimers localized in the same plane as mitochondria. Shown are Cy7.5, nuclei (Hoechst), mitochondria (MitoTracker Green), and merged images. The experiment was performed four times with similar results.

Article Snippet: T98G human GBM cells were obtained from the American Type Culture Collection (ATTC, Rockville, MD, USA) and cultured according to the provider instructions.

Techniques: Incubation, Labeling

Journal: Biomacromolecules

Article Title: Hydrolytically Stable Cationic Bis-MPA Dendrimers as Efficient Transfectants for Glioblastoma Cells and Primary Astrocytes

doi: 10.1021/acs.biomac.5c01202

Figure Lengend Snippet: Knockdown of p42-MAPK, Rheb, and MGMT proteins in T98G GBM cells. Dendriplexes formed by G3-CYS (1 μM) and different concentrations of siRNA (25 to 100 nM) targeting either (A) p42-MAPK, (B) Rheb, or (C) MGMT were incubated with T98G cells for 72 h. Cellular content was then quantified as described in the Experimental Procedures section. Data represent the mean, with error bars indicating the SEM of 4 to 6 experiments. * p < 0.05; * p < 0.001.

Article Snippet: T98G human GBM cells were obtained from the American Type Culture Collection (ATTC, Rockville, MD, USA) and cultured according to the provider instructions.

Techniques: Knockdown, Incubation