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human gbm cell lines a172  (ATCC)


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    ATCC human gbm cell lines a172
    Expression patterns of 14 representative CRRGs across the HEB, <t>A172,</t> SHG44, U251, and H4 cell lines. A – N qPCR analysis of CRRGs normalized to β-actin in HEB, A172, SHG44, U251, and H4 cell lines. ANOVA with Tukey's HSD test. All values are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs HEB. CRRGs expressions circadian rhythm-related genes, ANOVA analysis of variance
    Human Gbm Cell Lines A172, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human gbm cell lines a172/product/ATCC
    Average 86 stars, based on 1 article reviews
    human gbm cell lines a172 - by Bioz Stars, 2025-02
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    Images

    1) Product Images from "Circadian rhythm related genes signature in glioma for drug resistance prediction: a comprehensive analysis integrating transcriptomics and machine learning"

    Article Title: Circadian rhythm related genes signature in glioma for drug resistance prediction: a comprehensive analysis integrating transcriptomics and machine learning

    Journal: Discover Oncology

    doi: 10.1007/s12672-025-01863-2

    Expression patterns of 14 representative CRRGs across the HEB, A172, SHG44, U251, and H4 cell lines. A – N qPCR analysis of CRRGs normalized to β-actin in HEB, A172, SHG44, U251, and H4 cell lines. ANOVA with Tukey's HSD test. All values are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs HEB. CRRGs expressions circadian rhythm-related genes, ANOVA analysis of variance
    Figure Legend Snippet: Expression patterns of 14 representative CRRGs across the HEB, A172, SHG44, U251, and H4 cell lines. A – N qPCR analysis of CRRGs normalized to β-actin in HEB, A172, SHG44, U251, and H4 cell lines. ANOVA with Tukey's HSD test. All values are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs HEB. CRRGs expressions circadian rhythm-related genes, ANOVA analysis of variance

    Techniques Used: Expressing



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    (A–D) Relative lncRNA and mRNA expression in normal astrocyte (SVGP12) and <t>GBM</t> <t>cell</t> lines (U87, T98G, U343, LN229): (A) AC005229.4; (B) AC091182.2; (C) LIPT2; (D) GLS. (E) Tumor volumes were analyzed by bioluminescence imaging. (F) Hematoxylin and eosin staining of a representative GBM mouse brain (20×). (G-H) Relative mRNA expression in mouse GBM and normal brain tissues: (G) LIPT2; (H) GLS. * p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
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    Effect of Pazopanib on proliferation, apoptosis and migration in <t>GBM</t> tumor cell lines and HT-22 cells. ( A ) Cell proliferation were detected by CCK-8 in <t>lines-U87</t> and SW68, *** P < 0.001. ( B ) Cell apoptosis were detected by flow cytometry in U87 and SW68 cells, ** P < 0.01, *** P < 0.001. ( C ) Cell migration were detected in U87 and SW68 cells, ns P > 0.05. ( D ) Cell proliferation were detected by CCK-8 in HT-22 cells. ** P < 0.01. ( E ) Cell apoptosis were detected by flow cytometry in HT-22 cells under the co-intervention of RA and PAZ, *** P < 0.001. All experiments were repeated three times. Indicates a significant difference compared to other groups in a two-way ANOVA followed by Tukey’s test ( P < 0.05).
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    ( A ) Schematic diagram of the in vitro BBB monolayer model. ( B ) FL images of IVTPO, AOI987, and EB of in vitro BBB monolayer model. ( C ) Quantitative results of transcellular diffusion efficiency in (B). ( D ) Confocal images of macrophage cells and <t>U87-MG</t> cells after incubation with IVTPO (100 μg/ml) for 2 hours at 37°C. DAPI, 4′,6-diamidino-2-phenylindole. ( E ) Confocal images of tumor cells on designated time points after incubation with IVTPO (100 μg/ml). h, hours. ( F ) Cytotoxicity of different concentrations of IVTPO (10 to 100 μg/ml) to macrophage cells and U87-MG cells.
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    A Heatmap shows differentially expressed circRNAs between normoxia and hypoxia in <t>U87</t> cells on a scale from blue (downregulated, n = 2158) to red (upregulated, n = 2566). B Volcano plot shows differently expressed circRNAs between normoxia and hypoxia in U87 cells, n = 4727. Horizontal dashed line labels P = 0.05. Vertical dashed lines label |log2fold-change | = 1. C Verification of 17 up-regulated circRNAs ( P -value < 0.05 and reads counts >10) by qRT-PCR, n = 3. D Basal expression levels of 9 up-regulated circRNAs in U87 cells under hypoxia examined by qRT-PCR, n = 3. E Verification of corresponding circRNAs in U87 cells by RT-PCR using divergent and convergent primers. F Schematic illustration of the generation of circPLOD2a/b. G Junction sites of circPLOD2a/b were validated by Sanger sequencing. H Relative levels of circPLOD2a/b after treated with or without RNase R digesting determined by qRT-PCR in U87 cells, n = 3. I , J Relative expression levels of ( I ) circPLOD2a and ( J ) circPLOD2b under normoxia and hypoxia in <t>GBM</t> cell lines determined by qRT-PCR, n = 3. K Cellular localization of circPLOD2b examined by FISH with a junction specific antisense probe (red). Scale bar: 25 μm. L U87 cells were treated for 0 h, 12 h, 24 h, 48 h under 1% O 2 . Protein level of HIF1α and expression level of circPLOD2a/b were verified by western blot and qRT-PCR respectively, n = 3. M , N Protein level of HIF1α and expression level of circPLOD2a/b in ( M ) OE-HIF1α and ( N ) KD-HIF1α U87 cells under hypoxia, n = 3. Relative integrated density normalized to β-actin was marked above each band. Data are shown as mean ± SEM (error bars) and were analyzed using Student’s t -test ( M ) and ANOVA ( C – N ). NA, not available.
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    Image Search Results


    (A–D) Relative lncRNA and mRNA expression in normal astrocyte (SVGP12) and GBM cell lines (U87, T98G, U343, LN229): (A) AC005229.4; (B) AC091182.2; (C) LIPT2; (D) GLS. (E) Tumor volumes were analyzed by bioluminescence imaging. (F) Hematoxylin and eosin staining of a representative GBM mouse brain (20×). (G-H) Relative mRNA expression in mouse GBM and normal brain tissues: (G) LIPT2; (H) GLS. * p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Journal: PLOS ONE

    Article Title: Cuproptosis-related lncRNAs and genes: Potential markers for glioblastoma prognosis and treatment

    doi: 10.1371/journal.pone.0315927

    Figure Lengend Snippet: (A–D) Relative lncRNA and mRNA expression in normal astrocyte (SVGP12) and GBM cell lines (U87, T98G, U343, LN229): (A) AC005229.4; (B) AC091182.2; (C) LIPT2; (D) GLS. (E) Tumor volumes were analyzed by bioluminescence imaging. (F) Hematoxylin and eosin staining of a representative GBM mouse brain (20×). (G-H) Relative mRNA expression in mouse GBM and normal brain tissues: (G) LIPT2; (H) GLS. * p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Article Snippet: RNA from human GBM cell lines, the GBM cuproptosis cell model and the GBM mouse model was extracted with Trizol (Invitrogen, USA) according to the manufacturer’s protocol.

    Techniques: Expressing, Imaging, Staining

    (A) Western blot analysis of GBM cell lines treated with 10 μM CuCl 2 and 40 nM Elesclomol for 2 h. (B) Cell viability of the GBM cuproptosis model as evaluated by CCK-8 assay. (C-J) Quantitative real-time PCR analysis of AC005229.4, AC091182.2, LIPT2 and GLS in the GBM cuproptosis model. * p < 0.05, **p < 0.01, ***p < 0.001. ns, no significance.

    Journal: PLOS ONE

    Article Title: Cuproptosis-related lncRNAs and genes: Potential markers for glioblastoma prognosis and treatment

    doi: 10.1371/journal.pone.0315927

    Figure Lengend Snippet: (A) Western blot analysis of GBM cell lines treated with 10 μM CuCl 2 and 40 nM Elesclomol for 2 h. (B) Cell viability of the GBM cuproptosis model as evaluated by CCK-8 assay. (C-J) Quantitative real-time PCR analysis of AC005229.4, AC091182.2, LIPT2 and GLS in the GBM cuproptosis model. * p < 0.05, **p < 0.01, ***p < 0.001. ns, no significance.

    Article Snippet: RNA from human GBM cell lines, the GBM cuproptosis cell model and the GBM mouse model was extracted with Trizol (Invitrogen, USA) according to the manufacturer’s protocol.

    Techniques: Western Blot, CCK-8 Assay, Real-time Polymerase Chain Reaction

    Expression patterns of 14 representative CRRGs across the HEB, A172, SHG44, U251, and H4 cell lines. A – N qPCR analysis of CRRGs normalized to β-actin in HEB, A172, SHG44, U251, and H4 cell lines. ANOVA with Tukey's HSD test. All values are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs HEB. CRRGs expressions circadian rhythm-related genes, ANOVA analysis of variance

    Journal: Discover Oncology

    Article Title: Circadian rhythm related genes signature in glioma for drug resistance prediction: a comprehensive analysis integrating transcriptomics and machine learning

    doi: 10.1007/s12672-025-01863-2

    Figure Lengend Snippet: Expression patterns of 14 representative CRRGs across the HEB, A172, SHG44, U251, and H4 cell lines. A – N qPCR analysis of CRRGs normalized to β-actin in HEB, A172, SHG44, U251, and H4 cell lines. ANOVA with Tukey's HSD test. All values are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs HEB. CRRGs expressions circadian rhythm-related genes, ANOVA analysis of variance

    Article Snippet: The normal human brain glial cell line HEB and the human GBM cell lines A172, SHG44, U251, and H4 were procured from the American Type Culture Collection (ATCC; Manassas, USA).

    Techniques: Expressing

    Effect of Pazopanib on proliferation, apoptosis and migration in GBM tumor cell lines and HT-22 cells. ( A ) Cell proliferation were detected by CCK-8 in lines-U87 and SW68, *** P < 0.001. ( B ) Cell apoptosis were detected by flow cytometry in U87 and SW68 cells, ** P < 0.01, *** P < 0.001. ( C ) Cell migration were detected in U87 and SW68 cells, ns P > 0.05. ( D ) Cell proliferation were detected by CCK-8 in HT-22 cells. ** P < 0.01. ( E ) Cell apoptosis were detected by flow cytometry in HT-22 cells under the co-intervention of RA and PAZ, *** P < 0.001. All experiments were repeated three times. Indicates a significant difference compared to other groups in a two-way ANOVA followed by Tukey’s test ( P < 0.05).

    Journal: Scientific Reports

    Article Title: Genes related to neural tube defects and glioblastoma

    doi: 10.1038/s41598-025-86891-2

    Figure Lengend Snippet: Effect of Pazopanib on proliferation, apoptosis and migration in GBM tumor cell lines and HT-22 cells. ( A ) Cell proliferation were detected by CCK-8 in lines-U87 and SW68, *** P < 0.001. ( B ) Cell apoptosis were detected by flow cytometry in U87 and SW68 cells, ** P < 0.01, *** P < 0.001. ( C ) Cell migration were detected in U87 and SW68 cells, ns P > 0.05. ( D ) Cell proliferation were detected by CCK-8 in HT-22 cells. ** P < 0.01. ( E ) Cell apoptosis were detected by flow cytometry in HT-22 cells under the co-intervention of RA and PAZ, *** P < 0.001. All experiments were repeated three times. Indicates a significant difference compared to other groups in a two-way ANOVA followed by Tukey’s test ( P < 0.05).

    Article Snippet: Human GBM cell lines U87 and SWO-68 were obtained from the American Type Culture Collection ATCC (Manassas, VA, USA), and were cultured with the DMEM containing 10% fetal bovine serum (FBS, Invitrogen; Thermo Fisher Scientific, Inc. Waltham, MA, USA) at 37 ℃ with 5% CO 2 .

    Techniques: Migration, CCK-8 Assay, Flow Cytometry

    ( A ) Schematic diagram of the in vitro BBB monolayer model. ( B ) FL images of IVTPO, AOI987, and EB of in vitro BBB monolayer model. ( C ) Quantitative results of transcellular diffusion efficiency in (B). ( D ) Confocal images of macrophage cells and U87-MG cells after incubation with IVTPO (100 μg/ml) for 2 hours at 37°C. DAPI, 4′,6-diamidino-2-phenylindole. ( E ) Confocal images of tumor cells on designated time points after incubation with IVTPO (100 μg/ml). h, hours. ( F ) Cytotoxicity of different concentrations of IVTPO (10 to 100 μg/ml) to macrophage cells and U87-MG cells.

    Journal: Science Advances

    Article Title: Amphiphilic hemicyanine molecular probes crossing the blood-brain barrier for intracranial optical imaging of glioblastoma

    doi: 10.1126/sciadv.adq5816

    Figure Lengend Snippet: ( A ) Schematic diagram of the in vitro BBB monolayer model. ( B ) FL images of IVTPO, AOI987, and EB of in vitro BBB monolayer model. ( C ) Quantitative results of transcellular diffusion efficiency in (B). ( D ) Confocal images of macrophage cells and U87-MG cells after incubation with IVTPO (100 μg/ml) for 2 hours at 37°C. DAPI, 4′,6-diamidino-2-phenylindole. ( E ) Confocal images of tumor cells on designated time points after incubation with IVTPO (100 μg/ml). h, hours. ( F ) Cytotoxicity of different concentrations of IVTPO (10 to 100 μg/ml) to macrophage cells and U87-MG cells.

    Article Snippet: Orthotopic xenografting GBM model preparation: The U87-MG human GBM multiforme cell line was obtained from the American Type Culture Collection and maintained in Dulbecco’s modified Eagle’s medium (Gibco, 21063-029) supplemented with 10% FBS (Gibco, 10270-106), and 1% antibiotic-antimycotic solution at 37°C in a 5% CO 2 atmosphere.

    Techniques: In Vitro, Diffusion-based Assay, Incubation

    A Heatmap shows differentially expressed circRNAs between normoxia and hypoxia in U87 cells on a scale from blue (downregulated, n = 2158) to red (upregulated, n = 2566). B Volcano plot shows differently expressed circRNAs between normoxia and hypoxia in U87 cells, n = 4727. Horizontal dashed line labels P = 0.05. Vertical dashed lines label |log2fold-change | = 1. C Verification of 17 up-regulated circRNAs ( P -value < 0.05 and reads counts >10) by qRT-PCR, n = 3. D Basal expression levels of 9 up-regulated circRNAs in U87 cells under hypoxia examined by qRT-PCR, n = 3. E Verification of corresponding circRNAs in U87 cells by RT-PCR using divergent and convergent primers. F Schematic illustration of the generation of circPLOD2a/b. G Junction sites of circPLOD2a/b were validated by Sanger sequencing. H Relative levels of circPLOD2a/b after treated with or without RNase R digesting determined by qRT-PCR in U87 cells, n = 3. I , J Relative expression levels of ( I ) circPLOD2a and ( J ) circPLOD2b under normoxia and hypoxia in GBM cell lines determined by qRT-PCR, n = 3. K Cellular localization of circPLOD2b examined by FISH with a junction specific antisense probe (red). Scale bar: 25 μm. L U87 cells were treated for 0 h, 12 h, 24 h, 48 h under 1% O 2 . Protein level of HIF1α and expression level of circPLOD2a/b were verified by western blot and qRT-PCR respectively, n = 3. M , N Protein level of HIF1α and expression level of circPLOD2a/b in ( M ) OE-HIF1α and ( N ) KD-HIF1α U87 cells under hypoxia, n = 3. Relative integrated density normalized to β-actin was marked above each band. Data are shown as mean ± SEM (error bars) and were analyzed using Student’s t -test ( M ) and ANOVA ( C – N ). NA, not available.

    Journal: Communications Biology

    Article Title: Hypoxia-induced circPLOD2a/b promotes the aggressiveness of glioblastoma by suppressing XIRP1 through binding to HuR

    doi: 10.1038/s42003-025-07503-3

    Figure Lengend Snippet: A Heatmap shows differentially expressed circRNAs between normoxia and hypoxia in U87 cells on a scale from blue (downregulated, n = 2158) to red (upregulated, n = 2566). B Volcano plot shows differently expressed circRNAs between normoxia and hypoxia in U87 cells, n = 4727. Horizontal dashed line labels P = 0.05. Vertical dashed lines label |log2fold-change | = 1. C Verification of 17 up-regulated circRNAs ( P -value < 0.05 and reads counts >10) by qRT-PCR, n = 3. D Basal expression levels of 9 up-regulated circRNAs in U87 cells under hypoxia examined by qRT-PCR, n = 3. E Verification of corresponding circRNAs in U87 cells by RT-PCR using divergent and convergent primers. F Schematic illustration of the generation of circPLOD2a/b. G Junction sites of circPLOD2a/b were validated by Sanger sequencing. H Relative levels of circPLOD2a/b after treated with or without RNase R digesting determined by qRT-PCR in U87 cells, n = 3. I , J Relative expression levels of ( I ) circPLOD2a and ( J ) circPLOD2b under normoxia and hypoxia in GBM cell lines determined by qRT-PCR, n = 3. K Cellular localization of circPLOD2b examined by FISH with a junction specific antisense probe (red). Scale bar: 25 μm. L U87 cells were treated for 0 h, 12 h, 24 h, 48 h under 1% O 2 . Protein level of HIF1α and expression level of circPLOD2a/b were verified by western blot and qRT-PCR respectively, n = 3. M , N Protein level of HIF1α and expression level of circPLOD2a/b in ( M ) OE-HIF1α and ( N ) KD-HIF1α U87 cells under hypoxia, n = 3. Relative integrated density normalized to β-actin was marked above each band. Data are shown as mean ± SEM (error bars) and were analyzed using Student’s t -test ( M ) and ANOVA ( C – N ). NA, not available.

    Article Snippet: Human GBM cell lines (DBTRG, LN229, T98G, U251 and U87) and HEK 293 T were purchased from American Type Culture Collection (ATCC).

    Techniques: Quantitative RT-PCR, Expressing, Reverse Transcription Polymerase Chain Reaction, Sequencing, Western Blot

    A , B Results of transwell assays in ( A ) U87 and ( B ) LN229 OE-circPLOD2a cells under hypoxia. Scale bar: 100 μm. C , D Quantification of transwell assays in ( C ) U87 and ( D ) LN229 OE-circPLOD2a cells, n = 3. E , F Results of CCK8 assays on ( E ) U87 and ( F ) LN229 OE-circPLOD2a cells under hypoxia, n = 5. G , H Representative pictures and quantification of wound-healing assays in ( G ) U87 and ( H ) LN229 OE-circPLOD2a cells under hypoxia. Scale bar: 250 μm, n = 5. I , J Results of transwell assay in ( I ) U87 and ( J ) LN229 OE-circPLOD2b cells under hypoxia. Scale bar: 100 μm. K , L Quantification of transwell assays in ( K ) U87 and ( L ) LN229 OE-circPLOD2b cells, n = 3. M , N Results of CCK8 assay on ( M ) U87 and ( N ) LN229 OE-circPLOD2b cells, n = 5. O , P Representative pictures and quantification of wound-healing assay ( O ) U87 and ( P ) LN229 OE-circPLOD2b cells under hypoxia, n = 5. Scale bar: 250 μm. Data are shown as mean ± SEM (error bars) and analyzed using Student’s t- test ( C – P ) and ANOVA ( E – N ).

    Journal: Communications Biology

    Article Title: Hypoxia-induced circPLOD2a/b promotes the aggressiveness of glioblastoma by suppressing XIRP1 through binding to HuR

    doi: 10.1038/s42003-025-07503-3

    Figure Lengend Snippet: A , B Results of transwell assays in ( A ) U87 and ( B ) LN229 OE-circPLOD2a cells under hypoxia. Scale bar: 100 μm. C , D Quantification of transwell assays in ( C ) U87 and ( D ) LN229 OE-circPLOD2a cells, n = 3. E , F Results of CCK8 assays on ( E ) U87 and ( F ) LN229 OE-circPLOD2a cells under hypoxia, n = 5. G , H Representative pictures and quantification of wound-healing assays in ( G ) U87 and ( H ) LN229 OE-circPLOD2a cells under hypoxia. Scale bar: 250 μm, n = 5. I , J Results of transwell assay in ( I ) U87 and ( J ) LN229 OE-circPLOD2b cells under hypoxia. Scale bar: 100 μm. K , L Quantification of transwell assays in ( K ) U87 and ( L ) LN229 OE-circPLOD2b cells, n = 3. M , N Results of CCK8 assay on ( M ) U87 and ( N ) LN229 OE-circPLOD2b cells, n = 5. O , P Representative pictures and quantification of wound-healing assay ( O ) U87 and ( P ) LN229 OE-circPLOD2b cells under hypoxia, n = 5. Scale bar: 250 μm. Data are shown as mean ± SEM (error bars) and analyzed using Student’s t- test ( C – P ) and ANOVA ( E – N ).

    Article Snippet: Human GBM cell lines (DBTRG, LN229, T98G, U251 and U87) and HEK 293 T were purchased from American Type Culture Collection (ATCC).

    Techniques: Transwell Assay, CCK-8 Assay, Wound Healing Assay

    A , B Results of transwell assays in ( A ) U87 and ( B ) LN229 KD-circPLOD2a cells under hypoxia. Scale bar: 100 μm, n = 3. C , D Results of wound-healing assays in ( C ) U87 and ( D ) LN229 KD-circPLOD2a cells under hypoxia. Scale bar: 250 μm, n = 5. E , F Results of transwell assays in ( E ) U87 and ( F ) LN229 KD-circPLOD2b cells under hypoxia. Scale bar: 100 μm, n = 3. G , H Results of wound-healing assays in ( G ) U87 and ( H ) LN229 KD-circPLOD2b cells under hypoxia. Scale bar: 250 μm, n = 5. Data are shown as mean ± SEM (error bars) and analyzed using ANOVA. Scr: Scramble.

    Journal: Communications Biology

    Article Title: Hypoxia-induced circPLOD2a/b promotes the aggressiveness of glioblastoma by suppressing XIRP1 through binding to HuR

    doi: 10.1038/s42003-025-07503-3

    Figure Lengend Snippet: A , B Results of transwell assays in ( A ) U87 and ( B ) LN229 KD-circPLOD2a cells under hypoxia. Scale bar: 100 μm, n = 3. C , D Results of wound-healing assays in ( C ) U87 and ( D ) LN229 KD-circPLOD2a cells under hypoxia. Scale bar: 250 μm, n = 5. E , F Results of transwell assays in ( E ) U87 and ( F ) LN229 KD-circPLOD2b cells under hypoxia. Scale bar: 100 μm, n = 3. G , H Results of wound-healing assays in ( G ) U87 and ( H ) LN229 KD-circPLOD2b cells under hypoxia. Scale bar: 250 μm, n = 5. Data are shown as mean ± SEM (error bars) and analyzed using ANOVA. Scr: Scramble.

    Article Snippet: Human GBM cell lines (DBTRG, LN229, T98G, U251 and U87) and HEK 293 T were purchased from American Type Culture Collection (ATCC).

    Techniques:

    A – D Proteins pulled down by biotin-labeled circPLOD2a ( A ) or circPLOD2b ( C ) antisense probes and corresponding scrambled control probes in hypoxic U87 cells were visualized by SDS-PAGE and silver staining. Unique peptides identified by mass spectrometry in ( B ) circPLOD2a ( n = 73) or ( D ) circPLOD2b ( n = 54) compared with corresponding control lanes. E , F Potential proteins interacting with ( E ) circPLOD2a and ( F ) circPLOD2b were verified by RNA pull-down assays with corresponding antisense probes in hypoxic U87 cells. Relative integrated density normalized to β-actin was marked above each band. G – J RIP assay confirms the interaction of circPLOD2a/b and linear PLOD2 with HuR. CircPLOD2a/b and linear PLOD2 were detected by RT-PCR ( G ). The immunoprecipitation products of anti-HuR compared with IgG were treated with or without RNase R. Quantitative enrichment analysis of circPLOD2a ( H ), circPLOD2b ( I ) and linear PLOD2 ( J ) were analyzed by qRT-PCR, n = 3. K Cellular localization of circPLOD2b (red) and HuR (green) were examined by dual RNA-FISH and IF in U87 cells treated with normoxic and hypoxic conditions. Scale bar: 25 μm. L Schematic diagram of wild type and various truncated HuR with Flag-tag. (M) The interaction of circPLOD2a/b with full-length or truncated HuR protein were detected by RIP assay in U87 cells. CircPLOD2a/b were detected by RT-PCR; full-length and truncated HuR proteins were detected by western blot using anti-Flag antibody. N CircPLOD2a and circPLOD2b were immunoprecipitated by anti-Flag beads in U87 cells transfected with plasmids carrying HuR- wildtype (HuR-wt) or truncated forms (HuR-△RRM1/2/3) and detected by qRT-PCR, n = 3. Data are shown as mean ± SEM (error bars) and analyzed using Student’s t -test ( H – J ) and ANOVA ( N ).

    Journal: Communications Biology

    Article Title: Hypoxia-induced circPLOD2a/b promotes the aggressiveness of glioblastoma by suppressing XIRP1 through binding to HuR

    doi: 10.1038/s42003-025-07503-3

    Figure Lengend Snippet: A – D Proteins pulled down by biotin-labeled circPLOD2a ( A ) or circPLOD2b ( C ) antisense probes and corresponding scrambled control probes in hypoxic U87 cells were visualized by SDS-PAGE and silver staining. Unique peptides identified by mass spectrometry in ( B ) circPLOD2a ( n = 73) or ( D ) circPLOD2b ( n = 54) compared with corresponding control lanes. E , F Potential proteins interacting with ( E ) circPLOD2a and ( F ) circPLOD2b were verified by RNA pull-down assays with corresponding antisense probes in hypoxic U87 cells. Relative integrated density normalized to β-actin was marked above each band. G – J RIP assay confirms the interaction of circPLOD2a/b and linear PLOD2 with HuR. CircPLOD2a/b and linear PLOD2 were detected by RT-PCR ( G ). The immunoprecipitation products of anti-HuR compared with IgG were treated with or without RNase R. Quantitative enrichment analysis of circPLOD2a ( H ), circPLOD2b ( I ) and linear PLOD2 ( J ) were analyzed by qRT-PCR, n = 3. K Cellular localization of circPLOD2b (red) and HuR (green) were examined by dual RNA-FISH and IF in U87 cells treated with normoxic and hypoxic conditions. Scale bar: 25 μm. L Schematic diagram of wild type and various truncated HuR with Flag-tag. (M) The interaction of circPLOD2a/b with full-length or truncated HuR protein were detected by RIP assay in U87 cells. CircPLOD2a/b were detected by RT-PCR; full-length and truncated HuR proteins were detected by western blot using anti-Flag antibody. N CircPLOD2a and circPLOD2b were immunoprecipitated by anti-Flag beads in U87 cells transfected with plasmids carrying HuR- wildtype (HuR-wt) or truncated forms (HuR-△RRM1/2/3) and detected by qRT-PCR, n = 3. Data are shown as mean ± SEM (error bars) and analyzed using Student’s t -test ( H – J ) and ANOVA ( N ).

    Article Snippet: Human GBM cell lines (DBTRG, LN229, T98G, U251 and U87) and HEK 293 T were purchased from American Type Culture Collection (ATCC).

    Techniques: Labeling, Control, SDS Page, Silver Staining, Mass Spectrometry, Reverse Transcription Polymerase Chain Reaction, Immunoprecipitation, Quantitative RT-PCR, FLAG-tag, Western Blot, Transfection

    A Heatmap and ( B ) Volcano plot of DEGs in KD-circPLOD2a U87 cells compared to negative control cells under hypoxia. In total n = 57 up-regulated and n = 114 up-regulated genes were displayed. C , D Expression of 4 up-regulated genes in RNA-seq data in ( C ) KD-circPLOD2a and ( D ) OE-circPLOD2a U87 cells examined by qRT-PCR. E Expression of these 4 genes in KD-HuR cells. F , G Expression of XIRP1 in ( F ) KD-circPLOD2b and ( G ) OE-circPLOD2b cells. H , I Binding of ( H ) circPLOD2a and (I) XIRP1 to HuR were detected by RIP assay in U87 OE-circPLOD2a and control cells. J , K Binding of ( J ) circPLOD2b and ( K ) XIRP1 to HuR were detected by RIP assay in U87 OE-circPLOD2b and control cells. L , M The expression ( L ) mRNA and ( M ) protein level of XIRP1 were rescued by overexpression of HuR in OE-circPLOD2a and OE-circPLOD2b cells. Relative integrated density normalized to β-actin was marked above each band. N OE-HuR U87 and LN229 cells were treated with actinomycin D (5 μg/ml). The relative expression of XIRP1 was detected by qRT-PCR at different time points post actinomycin D treatment. O , P Transwell invasion assays illustrate that knockdown of XIRP1 in ( O ) KD-circPLOD2a or ( P ) KD-circPLOD2b U87 cells could rescue the inhibition of cell invasion and migration induced by knockdown of these two circRNAs under hypoxia. scale bar: 100 μm. (Q and R) Wound healing assays illustrate that knockdown of XIRP1 in ( Q ) KD-circPLOD2a or ( R ) KD-circPLOD2b U87 cells could rescue the inhibition of cell migration ability under hypoxia. Scale bar: 250 μm. n = 3 replicates unless otherwise noted. Data are shown as mean ± SEM (error bars) and analyzed using ANOVA. Scr: Scramble.

    Journal: Communications Biology

    Article Title: Hypoxia-induced circPLOD2a/b promotes the aggressiveness of glioblastoma by suppressing XIRP1 through binding to HuR

    doi: 10.1038/s42003-025-07503-3

    Figure Lengend Snippet: A Heatmap and ( B ) Volcano plot of DEGs in KD-circPLOD2a U87 cells compared to negative control cells under hypoxia. In total n = 57 up-regulated and n = 114 up-regulated genes were displayed. C , D Expression of 4 up-regulated genes in RNA-seq data in ( C ) KD-circPLOD2a and ( D ) OE-circPLOD2a U87 cells examined by qRT-PCR. E Expression of these 4 genes in KD-HuR cells. F , G Expression of XIRP1 in ( F ) KD-circPLOD2b and ( G ) OE-circPLOD2b cells. H , I Binding of ( H ) circPLOD2a and (I) XIRP1 to HuR were detected by RIP assay in U87 OE-circPLOD2a and control cells. J , K Binding of ( J ) circPLOD2b and ( K ) XIRP1 to HuR were detected by RIP assay in U87 OE-circPLOD2b and control cells. L , M The expression ( L ) mRNA and ( M ) protein level of XIRP1 were rescued by overexpression of HuR in OE-circPLOD2a and OE-circPLOD2b cells. Relative integrated density normalized to β-actin was marked above each band. N OE-HuR U87 and LN229 cells were treated with actinomycin D (5 μg/ml). The relative expression of XIRP1 was detected by qRT-PCR at different time points post actinomycin D treatment. O , P Transwell invasion assays illustrate that knockdown of XIRP1 in ( O ) KD-circPLOD2a or ( P ) KD-circPLOD2b U87 cells could rescue the inhibition of cell invasion and migration induced by knockdown of these two circRNAs under hypoxia. scale bar: 100 μm. (Q and R) Wound healing assays illustrate that knockdown of XIRP1 in ( Q ) KD-circPLOD2a or ( R ) KD-circPLOD2b U87 cells could rescue the inhibition of cell migration ability under hypoxia. Scale bar: 250 μm. n = 3 replicates unless otherwise noted. Data are shown as mean ± SEM (error bars) and analyzed using ANOVA. Scr: Scramble.

    Article Snippet: Human GBM cell lines (DBTRG, LN229, T98G, U251 and U87) and HEK 293 T were purchased from American Type Culture Collection (ATCC).

    Techniques: Negative Control, Expressing, RNA Sequencing Assay, Quantitative RT-PCR, Binding Assay, Control, Over Expression, Knockdown, Inhibition, Migration

    A , B ( A ) Representative bioluminescence images and ( B ) tumor growth curve of tumor-bearing mice intracranially transplanted with corresponding genetically modified U87 cells labeled with luciferase ( n = 5–6/ per group). C Relative expression of XIRP1 in CDX tumors, n = 3. D Representative images of IHC staining of Vimentin, E-cadherin and N-cadherin in CDX tumors. Scale bar: 100μm. E , F Relative protein level of Vimentin, E-cadherin, N-cadherin, HuR and XIRP1 in different genetically-modified ( E ) U87 cells in vitro and in ( F ) CDX tumors in vivo. Relative integrated density normalized to β-actin was marked above each band. Data are shown as mean ± SEM (error bars) and analyzed ANOVA. Scr: Scramble.

    Journal: Communications Biology

    Article Title: Hypoxia-induced circPLOD2a/b promotes the aggressiveness of glioblastoma by suppressing XIRP1 through binding to HuR

    doi: 10.1038/s42003-025-07503-3

    Figure Lengend Snippet: A , B ( A ) Representative bioluminescence images and ( B ) tumor growth curve of tumor-bearing mice intracranially transplanted with corresponding genetically modified U87 cells labeled with luciferase ( n = 5–6/ per group). C Relative expression of XIRP1 in CDX tumors, n = 3. D Representative images of IHC staining of Vimentin, E-cadherin and N-cadherin in CDX tumors. Scale bar: 100μm. E , F Relative protein level of Vimentin, E-cadherin, N-cadherin, HuR and XIRP1 in different genetically-modified ( E ) U87 cells in vitro and in ( F ) CDX tumors in vivo. Relative integrated density normalized to β-actin was marked above each band. Data are shown as mean ± SEM (error bars) and analyzed ANOVA. Scr: Scramble.

    Article Snippet: Human GBM cell lines (DBTRG, LN229, T98G, U251 and U87) and HEK 293 T were purchased from American Type Culture Collection (ATCC).

    Techniques: Genetically Modified, Labeling, Luciferase, Expressing, Immunohistochemistry, In Vitro, In Vivo

    A , B Relative expression of ( A ) circPLOD2a and ( B ) circPLOD2b were examined by qRT-PCR in clinical samples from patients with grade II-III ( n = 10) and grade IV ( n = 9) glioma. C Relative expression of XIRP1 in tissues from circPLOD2a-high and -low groups. The GBM tissues were divided into circPLOD2a-high ( n = 10) and -low ( n = 9) groups according to the expression of circPLOD2a. D Relative expression of XIRP1 in tissues from circPLOD2b-high and -low groups. The GBM tissues were divided into circPLOD2a-high ( n = 10) and -low ( n = 9) groups according to the expression of circPLOD2b. E Relative expression of XIRP1 in GBM and non-tumor brain tissues were analyzed from Rembrandt database. F Representative immunohistochemistry images of XIRP1 in a tissue microarray containing grade I ( n = 3), II ( n = 50), III ( n = 24) and IV ( n = 94) glioma. Scale bar: 100 μm. G , H Expression levels of XIRP1 in tissues from patients with different grade glioma were evaluated by ( G ) the percentage of cells with strong/moderate/weak intensity of XIRP1 staining and ( H ) histochemistry score (H-score) of each tissue on this microarray. I Kaplan–Meier survival curves of glioma patients in XIRP1-high and low groups. Glioma patients were categorized into XIRP1-high and -low group by the expression level of XIRP1 based on H-score. J Schematic illustration of HIF1α/circPLOD2a/b/HuR/XIRP1 axis. HIF1α induces the up-regulation of linear PLOD2 , circPLOD2a and circPLOD2b under hypoxia. CircPLOD2a/b suppress XIRP1 by competitively binding to HuR and enhance GBM cell aggressiveness. Data are shown as mean ± SEM (error bars) and analyzed using Student’s t -test ( A – E ), ANOVA ( H ) and Log-rank test ( I ).

    Journal: Communications Biology

    Article Title: Hypoxia-induced circPLOD2a/b promotes the aggressiveness of glioblastoma by suppressing XIRP1 through binding to HuR

    doi: 10.1038/s42003-025-07503-3

    Figure Lengend Snippet: A , B Relative expression of ( A ) circPLOD2a and ( B ) circPLOD2b were examined by qRT-PCR in clinical samples from patients with grade II-III ( n = 10) and grade IV ( n = 9) glioma. C Relative expression of XIRP1 in tissues from circPLOD2a-high and -low groups. The GBM tissues were divided into circPLOD2a-high ( n = 10) and -low ( n = 9) groups according to the expression of circPLOD2a. D Relative expression of XIRP1 in tissues from circPLOD2b-high and -low groups. The GBM tissues were divided into circPLOD2a-high ( n = 10) and -low ( n = 9) groups according to the expression of circPLOD2b. E Relative expression of XIRP1 in GBM and non-tumor brain tissues were analyzed from Rembrandt database. F Representative immunohistochemistry images of XIRP1 in a tissue microarray containing grade I ( n = 3), II ( n = 50), III ( n = 24) and IV ( n = 94) glioma. Scale bar: 100 μm. G , H Expression levels of XIRP1 in tissues from patients with different grade glioma were evaluated by ( G ) the percentage of cells with strong/moderate/weak intensity of XIRP1 staining and ( H ) histochemistry score (H-score) of each tissue on this microarray. I Kaplan–Meier survival curves of glioma patients in XIRP1-high and low groups. Glioma patients were categorized into XIRP1-high and -low group by the expression level of XIRP1 based on H-score. J Schematic illustration of HIF1α/circPLOD2a/b/HuR/XIRP1 axis. HIF1α induces the up-regulation of linear PLOD2 , circPLOD2a and circPLOD2b under hypoxia. CircPLOD2a/b suppress XIRP1 by competitively binding to HuR and enhance GBM cell aggressiveness. Data are shown as mean ± SEM (error bars) and analyzed using Student’s t -test ( A – E ), ANOVA ( H ) and Log-rank test ( I ).

    Article Snippet: Human GBM cell lines (DBTRG, LN229, T98G, U251 and U87) and HEK 293 T were purchased from American Type Culture Collection (ATCC).

    Techniques: Expressing, Quantitative RT-PCR, Immunohistochemistry, Microarray, Staining, Binding Assay