Journal: Cell Reports Medicine

Article Title: PPRX-1701, a nanoparticle formulation of 6′-bromoindirubin acetoxime, improves delivery and shows efficacy in preclinical GBM models

doi: 10.1016/j.xcrm.2023.101019

Figure Lengend Snippet:

Article Snippet: Mouse: GL261fluc2 , PerkinElmer , BW134246.

Techniques: Recombinant, Protease Inhibitor, Plasmid Preparation, Staining, cDNA Synthesis, SYBR Green Assay, Cell Isolation, Microarray, Software, Imaging, Light Microscopy

Journal: Molecular Therapy. Nucleic Acids

Article Title: Palmitoyl transferases act as potential regulators of tumor-infiltrating immune cells and glioma progression

doi: 10.1016/j.omtn.2022.04.030

Figure Lengend Snippet: Impact of 2-BP on glioma-cell viability and apoptosis (A) Gl261 cells were treated with 50 μM 2-BP for indicated times, and cell viability was measured (∗∗p < 0.01; not significant [ns], p > 0.05 compared with the control group). (B) Gl261 cells were treated with 50 μM 2-BP for indicated times, and effects of 2-BP on GL261 apoptotic rate (Q2+Q3) at 0 h (6.10%), 24 h (10.06%), and 48 h (19.54%) were evaluated. (C) C6 cells were treated with 50 μM 2-BP for indicated times, and cell viability was measured (ns; p > 0.05 compared with the control group). (D) C6 cells were treated with 50 μM 2-BP for indicated times, and effects of 2-BP on C6 apoptotic rate (Q2+Q3) at 0 h (3.93%), 24 h (3.40%), and 48 h (5.12%) were evaluated.

Article Snippet: The GL261 glioma cell line was kindly donated by Professor Jie Luo (Taihe Hospital, Hubei Province, China).

Techniques: Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: Palmitoyl transferases act as potential regulators of tumor-infiltrating immune cells and glioma progression

doi: 10.1016/j.omtn.2022.04.030

Figure Lengend Snippet: 2-BP inhibits glioma-cell autophagy (A) Autophagy-related key proteins of GL261 and C6 cells were detected by Western blotting assay. (B–D) Relative Beclin-1,P62, LC3B protein levels between control and 2-BP treated groups in GL261 and C6 cells. ∗p < 0.05; ∗∗p < 0.01; ns; p > 0.05 compared with the control group

Article Snippet: The GL261 glioma cell line was kindly donated by Professor Jie Luo (Taihe Hospital, Hubei Province, China).

Techniques: Western Blot, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: Palmitoyl transferases act as potential regulators of tumor-infiltrating immune cells and glioma progression

doi: 10.1016/j.omtn.2022.04.030

Figure Lengend Snippet: 2-BP promotes the sensitivity of glioma cells to TMZ chemotherapy (A) Gl261 cells were treated with 50 μM 2-BP and 200 μM TMZ for 48 h, and cell viability was measured (∗∗p < 0.01, compared with the control group; ## p < 0.01, compared with TMZ). (B) Gl261 cells were treated with 50 μM 2-BP and 200 μM TMZ for 48 h, and effects of 2-BP and TMZ on GL261 apoptotic rate (Q2+Q3) at 48 h were evaluated: control group (5.82%), TMZ (12.01%), and 2-BP + TMZ (38.90%). (C) C6 cells were treated with 50 μM 2-BP and 200 μM TMZ for 48 h, and cell viability was measured (ns; p > 0.05, compared with the control group; # p < 0.05, compared with TMZ). (D) C6 cells were treated with 50 μM 2-BP and 200 μM TMZ for 48 h, and effects of 2-BP and TMZ on C6 apoptotic rate (Q2+Q3) at 48 h were evaluated: control group (3.99%), TMZ (7.50%), and 2-BP + TMZ (6.70%).

Article Snippet: The GL261 glioma cell line was kindly donated by Professor Jie Luo (Taihe Hospital, Hubei Province, China).

Techniques: Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: Palmitoyl transferases act as potential regulators of tumor-infiltrating immune cells and glioma progression

doi: 10.1016/j.omtn.2022.04.030

Figure Lengend Snippet: Effects of 2-BP on glioma-induced microglia migration (A) Glioma cells were pretreated with 2-BP for 48 h and then co-cultured with BV2 cells. Effects of 2-BP-treated GL261 and C6 cells on BV2 cell migration were evaluated. (B and C) Glioma cells were treated with 2-BP for 48 h, and CCL2 and CXCL16 expression was detected by qRT-PCR (∗p < 0.05; ns; p > 0.05; compared with the control group). (D) Effects of 2-BP on microglia migration in vivo are shown.

Article Snippet: The GL261 glioma cell line was kindly donated by Professor Jie Luo (Taihe Hospital, Hubei Province, China).

Techniques: Migration, Cell Culture, Expressing, Quantitative RT-PCR, Control, In Vivo

Journal: ACS chemical biology

Article Title: Novel Imidazotetrazine Evades Known Resistance Mechanisms and Is Effective against Temozolomide-Resistant Brain Cancer in Cell Culture

doi: 10.1021/acschembio.2c00022

Figure Lengend Snippet: Investigation of the mechanism of action and efficacy of CPZ. (A) Detection of O6-methyl-2′-deoxyguanosine or O6-propargyl-2′-deoxyguanosine in GL261 cells after treatment with TMZ or CPZ, respectively. GL261 cells were treated with the indicated concentration of compound for 8 h before they were harvested and genomic DNA was extracted. DNA (10 μg) was hydrolyzed and submitted to LC–MS/MS for quantitation. Both O6-Me-dG and O6-proparyl-dG were below the limit of detection in the DMSO control and CPZ samples. Error is SEM, n ≥ 3.(B) Cytotoxicity of TMZ (left) or CPZ (right) in T98G cells pretreated (3 h) with MGMT inhibitor O6BG. Error is SEM, n ≥ 3. (C) Western blot of GL261, GL261 MGMT+, and T98G (10 μg protein loading, 1:1000 anti-MGMT antibody). (D) 7 day dose–response curves of TMZ and CPZ in GL261 cells. Error is SEM, n ≥ 3. (E) 7 day dose–response curves of TMZ and CPZ in GL261 MGMT+ cells. Error is SEM, n ≥ 3.

Article Snippet: Stable cell line generation of GL261 MGMT+ was purchased from Creative Biogene.

Techniques: Concentration Assay, Liquid Chromatography with Mass Spectroscopy, Quantitation Assay, Control, Western Blot

Journal: Materials Today Bio

Article Title: Delivery of extracellular vesicles loaded with immune checkpoint inhibitors for immunotherapeutic management of glioma

doi: 10.1016/j.mtbio.2024.101244

Figure Lengend Snippet: Barrier permeation enhancement effect of EVs and efficient delivery of aPD-L1/EV into brain. ( A ) Scheme of the in vitro barrier permeation test with C2BBe1 trans-well system (created from Biorender.com ). ( B ) Amounts of permeated aPD-L1 with free aPD-L1 or aPD-L1/EV administration through the C2BBe1 barrier. ( C ) Comparison of drug delivery efficiency with RO or IN administration of FITC-conjugated IgG or IgG/EV. The upper column displays the fluorescence of IgG within brain sections, and the lower column shows the cumulative fluorescence intensity of brain sections. ( D ) Distribution of aPD-L1 in GL261 tumor-bearing mice. Green fluorescence indicates aPD-L1, and nuclei were stained with DAPI. Statistics in ( B, C ) were conducted by one-way ANOVA with Bonferroni post hoc test. Asterisks represent a significant difference between indicated groups, with significance defined as p < 0.05. Data were presented as mean ± SD with n = 3.

Article Snippet: The GL261-luc murine glioblastoma cell line was purchased from Creative Biolabs (Shirley, NY, USA).

Techniques: In Vitro, Comparison, Fluorescence, Staining

Journal: Materials Today Bio

Article Title: Delivery of extracellular vesicles loaded with immune checkpoint inhibitors for immunotherapeutic management of glioma

doi: 10.1016/j.mtbio.2024.101244

Figure Lengend Snippet: Therapeutic effect of ICI/EVs in managing orthotopic GL261 tumors. ( A ) Schematic of experimental procedures for GL261 tumor management (created from Biorender.com ). ( B, C ) Tumor volumes were evaluated with IVIS, and representative images are shown in ( B ). Changes in tumor volume were calculated based on the total flux observed on day 0 ( C ). ( D ) The survival of orthotopic GL261 tumor-bearing mice post-treatment was analyzed. The aPD-L1/EV + aCTLA-4/EV group demonstrated an extended survival time compared to other treatment groups. ( E ) Ki67 staining of GL261 tumors. Scale bar: 50 μm. Statistics for ( C ) were obtained from GLM with Bonferroni post hoc test, and for ( D ) from the Kaplan-Meier survival analysis with log rank test. The asterisk in ( C ) indicates a significant difference from the PBS and aPD-L1 treatments. In ( D ), the asterisk and hash signs represent significant difference from all treatment groups and from the PBS group, respectively. Significance was defined as p < 0.05. Data are presented as mean ± SEM with n = 4 in ( C ) and in ( D ) with n = 8∼18.

Article Snippet: The GL261-luc murine glioblastoma cell line was purchased from Creative Biolabs (Shirley, NY, USA).

Techniques: Staining

Journal: Materials Today Bio

Article Title: Delivery of extracellular vesicles loaded with immune checkpoint inhibitors for immunotherapeutic management of glioma

doi: 10.1016/j.mtbio.2024.101244

Figure Lengend Snippet: Immune cells in GL261 tumors post treatment. ( A ) IHC images of cytotoxic T cells within the tumor tissue. Green fluorescence indicates CD8 expression on T cells, while nuclei are counterstained with DAPI. Scale bar: 50 μm. ( B ) IHC images of M1 macrophages within tumor tissue. Green fluorescence indicates F4/80-stained macrophages, and red fluorescence indicates iNOS expression. Nuclei are counterstained with DAPI. Scale bar: 20 μm. ( C-E ) Flow cytometry analysis results for total T cells ( C ), cytotoxic T cells ( D ), and M1 macrophages ( E ). Statistics for ( C, D, E ) were obtained from one-way ANOVA with Bonferroni post hoc test, with significance defined as p < 0.05. Data are expressed as mean ± SEM with n = 5.

Article Snippet: The GL261-luc murine glioblastoma cell line was purchased from Creative Biolabs (Shirley, NY, USA).

Techniques: Fluorescence, Expressing, Staining, Flow Cytometry