human gbm cell lines t98g  (Procell Inc)

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: bioRxiv

Article Title: Loss of PIMREG impairs double-strand break signaling and repair

doi: 10.1101/2025.10.15.682590

Figure Lengend Snippet: ( A-D ) Volcano plot graphs of PIMREG interacting proteins in U87MG (A, B) and T98G (C, D) cells. ( B, D ) TMZ-regulated PIMREG interactors (TMZ vs. DMSO). Upregulated proteins (red dots) interact with PIMREG only under TMZ treatment or show increased association under treatment. Downregulated proteins (blue dots) exhibit reduced or lost binding upon TMZ treatment. ( A, C ) PIMREG interacting proteins under vehicle (DMSO) conditions (PIMREG IP vs. IgG). Proteins with adjusted p -value < 0.05 and log 2 fold change > 1 (PIMREG IP vs. IgG) or log 2 fold change < -1 and >1 (TMZ vs. DMSO) were considered significant (highlighted as blue or red dots). ( E ) Interaction network of proteins identified as PIMREG interactors in U87MG cells (PIMREG IP vs. IgG). ( F ) Interaction network of the proteins identified as TMZ-regulated PIMREG interactors in U87MG and ( G ) in T98G cells. Each node represents one protein. The proteins interacting with each other are represented in clusters. Node colors indicate proteins that have the same function within the clusters. Lines connecting the nodes represent known interactions (cyan: from curated databases; magenta: experimentally determined), predicted interactions (green: gene neighborhood; red: gene fusions; blue: gene co-occurrence) and other types of interactions (black: co-expression; yellow: text mining; light blue: protein homology). ( H, I, J ) Biological processes (BP) in each group of PIMREG interacting proteins. Number of ontological terms grouped as a BP (graph bars) is depicted in the x axis. Number of genes involved in each BP is shown next to each bar. The bars in gray represent previously described functions of PIMREG. The hatched bars represent the putative new functions of PIMREG.

Article Snippet: The GBM human cell line T98G was obtained from the American Type Culture Collection (ATCC, Manassas, USA) and U87MG from the Banco de Células do Rio de Janeiro (BCRJ, Rio de Janeiro, Brazil).

Techniques: Binding Assay, Expressing

Journal: bioRxiv

Article Title: Loss of PIMREG impairs double-strand break signaling and repair

doi: 10.1101/2025.10.15.682590

Figure Lengend Snippet: Lysates of T98G cells untreated (UT) or treated with TMZ and vehicle (DMSO) for 24 h were precipitated with ( A ) anti-PIMREG antibody and isotype control (IgG). Western blot analysis with anti-PIMREG antibody shows the precipitated protein in the upper panel. Precipitated PIMREG is indicated with the arrowhead (lanes 3, 6 and 9). Immunoblotting of the precipitated protein with anti-phospho-ATM/ATR substrate antibody is shown in the lower panel. Asterisks (*) indicate the precipitated PIMREG protein detected with the anti-phospho-ATM/ATR substrate antibody. ( B ) ATM/ATR substrates immunoprecipitated using anti-phospho-ATM/ATR substrate antibody were immunoblotted with the same antibody (upper panel) and anti-PIMREG antibody (lower panel). Asterisk (*) indicates the precipitated phospho-ATM/ATR substrates detected with anti-PIMREG antibody (lane 9). As input control, lanes 1, 4 and 7 were loaded with 20 % of the protein extract used for each precipitation reaction. UT: untreated.

Article Snippet: The GBM human cell line T98G was obtained from the American Type Culture Collection (ATCC, Manassas, USA) and U87MG from the Banco de Células do Rio de Janeiro (BCRJ, Rio de Janeiro, Brazil).

Techniques: Control, Western Blot, Immunoprecipitation

Journal: bioRxiv

Article Title: Loss of PIMREG impairs double-strand break signaling and repair

doi: 10.1101/2025.10.15.682590

Figure Lengend Snippet: ( A ) Western blot analysis of protein extracts from T98G WT and PIMREG KO (KO-1, KO-2, KO-3 and KO-4) cells. Membranes were blotted with anti-PIMREG, anti-phospho-H2AX (S139) antibodies. Actin was used as a loading control. ( B ) Bar graph showing densitometric quantification of γH2AX bands, normalized to their corresponding endogenous actin levels (γH2AX/actin). Results are shown as mean ± SD (standard deviation) of three independent experiments. Red dashed line indicates γH2AX levels in WT cells for comparison. ( C ) Western blot analysis of protein extracts derived from T98G WT and PIMREG KOs treated with TMZ or vehicle (DMSO) for 24 h. ( D ) Bar graph showing γH2AX accumulation expressed as the ratio of γH2AX levels (γH2AX/actin) in TMZ-treated cells relative to their corresponding DMSO control (TMZ/DMSO). Results are shown as mean ± SD (standard deviation) of five independent experiments. Red dashed line indicates the γH2AX accumulation in WT cells upon TMZ treatment. ( E ) Western blot analysis of protein extracts from T98G WT and PIMREG KO cells following IR exposure. Samples were collected at 0.5 and 1h post-irradiation. Tubulin was used as a loading control. ( F ) Bar graph showing γH2AX accumulation expressed as the ratio of γH2AX levels (γH2AX/tubulin) in IR-treated cells relative to their corresponding non-irradiated controls (IR/NI). Results are shown as mean ± SD (standard deviation) of four independent experiments. Red dashed line indicates γH2AX accumulation in WT cells following IR exposure. Representative western blots are shown. (*) p -value < 0.05, One-way ANOVA test.

Article Snippet: The GBM human cell line T98G was obtained from the American Type Culture Collection (ATCC, Manassas, USA) and U87MG from the Banco de Células do Rio de Janeiro (BCRJ, Rio de Janeiro, Brazil).

Techniques: Western Blot, Control, Standard Deviation, Comparison, Derivative Assay, Irradiation

Journal: bioRxiv

Article Title: Loss of PIMREG impairs double-strand break signaling and repair

doi: 10.1101/2025.10.15.682590

Figure Lengend Snippet: Representative immunofluorescence analysis of T98G WT and PIMREG KO cells treated with ( A ) TMZ or vehicle (DMSO) for 24 h and ( C ) exposed to IR (4 Gy) and collected at 0.5 h post-irradiation. PIMREG is shown in green (Alexa Fluor 488), γH2AX in red (Alexa Fluor 555) and nuclei (DAPI) in cyan. Scale bar: 10 µM. ( B, D ) Fluorescence intensity quantification of nuclear γH2AX, quantified in at least 75 cells per condition. Data are from 3 to 5 independent experiments. The horizontal bars indicate the median intensity. (**) p -value < 0.01, (***) p -value < 0.001 and (****) p -value < 0.0001, One-way ANOVA test followed by the post-test Kruskal-Wallis was applied. WT: wild-type; KO: knockout; KO-2-recPIMREG: KO-2 cells that recovered PIMREG’s expression; NI: non-irradiated. ( E ) Western blot analysis of PIMREG in the same samples in (C and D): T98G WT, PIMREG KOs (KO-1 and KO-3) and KO-2-recPIMREG. Membranes were blotted with anti-PIMREG and actin was used as a loading control.

Article Snippet: The GBM human cell line T98G was obtained from the American Type Culture Collection (ATCC, Manassas, USA) and U87MG from the Banco de Células do Rio de Janeiro (BCRJ, Rio de Janeiro, Brazil).

Techniques: Immunofluorescence, Irradiation, Fluorescence, Knock-Out, Expressing, Western Blot, Control

Journal: bioRxiv

Article Title: Loss of PIMREG impairs double-strand break signaling and repair

doi: 10.1101/2025.10.15.682590

Figure Lengend Snippet: ( A, C, E ) Western blot analysis of protein extracts derived from T98G WT and PIMREG KOs treated with TMZ or vehicle (DMSO) for 24 h. Membranes were blotted with anti-Phospho-KAP1 (Ser824) and KAP1 (A), anti-Phospho CHK1 (Ser345) and CHK1 (C) and anti-Phospho-DNA-PKc (Ser2056) and DNA-PKc (E) antibodies. Tubulin was used as a loading control. ( B, D, F ) Bar graph showing densitometric quantification of phosphorylated protein band intensities normalized to total protein levels. Protein activation is expressed as the ratio of the phosphorylation levels in TMZ-treated cells relative to their corresponding DMSO control (TMZ/DMSO). Results are shown as mean ± SD (standard deviation) of three independent experiment. Red dashed line indicates the protein activation in WT cells upon TMZ treatment. ( G, I, K, M ) Western blot analysis of protein extracts from T98G WT and PIMREG KO cells (KO-1, KO-2, KO-3, and KO-4) following exposure to IR (4Gy). Samples were collected at 0.5 and 1h post-irradiation. Membranes were blotted with anti-Phospho ATM (Ser1981) and ATM (G), anti-Phospho-KAP1 (Ser824) and KAP1 (I), anti-Phospho CHK1 (Ser345) and CHK1 (K), anti-Phospho-DNA-PKc (Ser2056) and DNA-PKc (M) antibodies. Tubulin was used as a loading control. ( H, J, L, N ) Bar graph showing densitometric quantification of phosphorylated protein band intensities normalized to total protein levels. Protein activation is expressed as the ratio of the phosphorylation levels in IR samples relative to their corresponding non-irradiated controls (IR/NI). Results are shown as mean ± SD (standard deviation) of four independent experiment. Red dashed line indicates the protein activation in WT cells at the indicated time points following IR exposure. (*) p -value < 0.05 and (**) p -value < 0.01, One-way ANOVA test. WT: wild-type; KO: knockout; NI: non-irradiated.

Article Snippet: The GBM human cell line T98G was obtained from the American Type Culture Collection (ATCC, Manassas, USA) and U87MG from the Banco de Células do Rio de Janeiro (BCRJ, Rio de Janeiro, Brazil).

Techniques: Western Blot, Derivative Assay, Control, Activation Assay, Phospho-proteomics, Standard Deviation, Irradiation, Knock-Out