Journal: Bioengineering & Translational Medicine

Article Title: Utilizing induced neural stem cell‐based delivery of a cytokine cocktail to enhance chimeric antigen receptor‐modified T‐cell therapy for brain cancer

doi: 10.1002/btm2.10538

Figure Lengend Snippet: RANTES/IL‐15‐secreting GBM tumor cells enhance T‐cell trafficking and expansion. (a) RANTES and IL‐15 were detected in culture supernatant collected from transduced U87 tumor cells. (b) U87 or U87 RANTES‐IL‐15 were seeded and cultured for 24 or 48 h. Transwell inserts were added and CSPG4‐CAR‐T or control T cells were seeded therein. T cells were allowed to migrate through inserts for 5 h. Afterward, inserts were removed and media was collected for flow cytometry. (c) Migration of CSPG4‐CAR‐T or control T cells toward U87 or U87 RANTES‐IL‐15 cultured for 24 or 48 h was evaluated by flow cytometry ( n = 3). (d) NSG mice were implanted i.c. with U87, U87 RANTES‐IL‐15 , or PBS in the right hemisphere of the brain. Five days later, mice were infused i.v. with CSPG4‐CAR‐T. Mice were sacrificed 1, 4, or 7 days following CSPG4‐CAR‐T infusion. Brains were harvested, bisected along the interhemispheric fissure, and processed for flow cytometry ( n = 3). (e) CSPG4‐CAR‐T concentrations in single cell suspensions generated from bisected mouse brains (contralateral hemisphere vs tumor hemisphere) were analyzed by flow cytometry. Schematic(s) created with BioRender.com . * p < 0.05, ** p < 0.01, *** p < 0.001. CAR‐T, chimeric antigen receptor‐modified T cell; CSPG4‐CAR‐T, CAR‐T cells targeting chondroitin sulfate proteoglycan 4 antigen; GBM, glioblastoma; IL‐15, interuekin 15; NSG, NOD.Cg‐ Prkdc scid Il2rg tm1Wjl /SzJ.

Article Snippet: The U87 glioma cell line and human telomerase reverse transcriptase (hTERT)‐immortalized normal human fibroblasts (NHF1) were obtained from the American Type Culture Collection and as previously described.

Techniques: Cell Culture, Control, Flow Cytometry, Migration, Generated, Modification

Journal: Bioengineering & Translational Medicine

Article Title: Utilizing induced neural stem cell‐based delivery of a cytokine cocktail to enhance chimeric antigen receptor‐modified T‐cell therapy for brain cancer

doi: 10.1002/btm2.10538

Figure Lengend Snippet: Development and characterization of hiNeuroS RANTES‐IL‐15 . (a) Schematic representation of lentivirus encoding human RANTES and IL‐15. (b) Human fibroblasts were transduced with LV‐RANTES‐IL‐15‐GFP, LV‐Sox2, and LV‐rtTA in succession. Reprogramming and primary hiNeuroS formation were initiated by switching to NSC growth media supplemented with growth factors and tetracycline. Three rounds of cell cluster selection were performed by serial seeding on laminin‐coated plates. After each round of selection, nonadhering cells were collected and expanded to yield secondary hiNeuroS. (c) Fluorescence imaging of hiNeuroS RANTES‐IL‐15 confirms successful LV‐RANTES‐IL‐15‐GFP transduction. Scale bar represents 100 μm. (d) RANTES and IL‐15 were detected in culture supernatant collected from transduced hiNeuroS. (e) Migration of CSPG4‐CAR‐T toward U87 ± NS‐CM or control media using a transwell migration assay was evaluated by flow cytometry ( n = 3). (f) CSPG4‐CAR‐T counts after 3 days in co‐culture with U87 ± NS‐CM ( n = 3). (g) Anti‐tumor activity of CSPG4‐CAR‐T after 3 days in co‐culture with U87 ± NS‐CM was assessed by bioluminescence ( n = 8–10). Schematic(s) created with BioRender.com . * p < 0.05, ** p < 0.01, *** p < 0.001. CAR‐T, chimeric antigen receptor‐modified T cell; CSPG4‐CAR‐T, CAR‐T cells targeting chondroitin sulfate proteoglycan 4 antigen; NSC, neural stem cell.

Article Snippet: The U87 glioma cell line and human telomerase reverse transcriptase (hTERT)‐immortalized normal human fibroblasts (NHF1) were obtained from the American Type Culture Collection and as previously described.

Techniques: Transduction, Selection, Fluorescence, Imaging, Migration, Control, Transwell Migration Assay, Flow Cytometry, Co-Culture Assay, Activity Assay, Modification

Journal: Bioengineering & Translational Medicine

Article Title: Utilizing induced neural stem cell‐based delivery of a cytokine cocktail to enhance chimeric antigen receptor‐modified T‐cell therapy for brain cancer

doi: 10.1002/btm2.10538

Figure Lengend Snippet: Combination hiNeuroS RANTES‐IL‐15 + CAR‐T therapy increases CAR‐T infiltration and eradicates non‐invasive GBM. (a) Schematic representation of study design. NSG mice were implanted i.c. with FLuc + U87 ± hiNeuroS RANTES‐IL‐15 in the right hemisphere of the brain. Four days later, mice were infused i.v. with CSPG4‐CAR‐T or PBS ( n = 5–7). (b) Representative bioluminescent images depict U87 tumor growth. (c) Luminescence was quantified by calculating total flux within a region of interest centered on the head, then normalizing to Day 0 values. Dotted line signifies CAR‐T infusion. (d) Kaplan–Meier survival curves. (e) AUC was calculated for each normalized tumor growth curve. (f) CAR‐T tumor infiltration (brown) was assessed by immunohistochemistry. Schematic(s) created with BioRender.com . *** p < 0.001. CAR‐T, chimeric antigen receptor‐modified T cell; CSPG4‐CAR‐T, CAR‐T cells targeting chondroitin sulfate proteoglycan 4 antigen; GBM, glioblastoma; NSG, NOD.Cg‐ Prkdc scid Il2rg tm1Wjl /SzJ.

Article Snippet: The U87 glioma cell line and human telomerase reverse transcriptase (hTERT)‐immortalized normal human fibroblasts (NHF1) were obtained from the American Type Culture Collection and as previously described.

Techniques: Immunohistochemistry, Modification

Journal: Amino Acids

Article Title: Arginine deprivation affects glioblastoma cell adhesion, invasiveness and actin cytoskeleton organization by impairment of β-actin arginylation

doi: 10.1007/s00726-014-1857-1

Figure Lengend Snippet: Effect of arginine deprivation on glioblastoma cells. a Cell growth of U251 ( upper panel ) and U87 ( lower panel ) cells cultivated in control, -Arg, -Lys and conditions. b U251 cell viability assessed under deprivation and re-supplementation conditions, as indicated. Upper and lower panels , arginine and lysine re-supplementation, respectively. 100 %, the number of the viable cells at time 0. Data in a and b are means ± SD; *** and *Statistical relevance p < 0.001 and p < 0.05, respectively

Article Snippet: Human glioblastoma U251 MG (U251) and U87 MG (U87) cell lines were purchased from CLS Cell Lines Service (Germany) and monitored for correct genetic profile via microsatellite analyses according to a protocol described earlier (Peickert et al. ).

Techniques:

Journal: Amino Acids

Article Title: Arginine deprivation affects glioblastoma cell adhesion, invasiveness and actin cytoskeleton organization by impairment of β-actin arginylation

doi: 10.1007/s00726-014-1857-1

Figure Lengend Snippet: Arginine deprivation affects morphology of glioblastoma but not glia cells. a , b , d Rat glia, U251 and U87 cells stained with Alexa 488-phalloidin, respectively. c Micrographs of U251 cells attained with scanning electron microscope. Insets in a , b and d ~2–3× magnification of the marked areas . Bars , in a , b and d 50 μm, and in c 10 μm. e U251 cells stained with Alexa 488-phalloidin before and after re-supplementation with Arg or Lys up to 0.4 and 0.8 mM concentration, respectively. Arrows point to lamellipodia, arrowheads point to elongated cells

Article Snippet: Human glioblastoma U251 MG (U251) and U87 MG (U87) cell lines were purchased from CLS Cell Lines Service (Germany) and monitored for correct genetic profile via microsatellite analyses according to a protocol described earlier (Peickert et al. ).

Techniques: Staining, Microscopy, Concentration Assay

Journal: Amino Acids

Article Title: Arginine deprivation affects glioblastoma cell adhesion, invasiveness and actin cytoskeleton organization by impairment of β-actin arginylation

doi: 10.1007/s00726-014-1857-1

Figure Lengend Snippet: Arginine deprivation impairs cell motility. a , b Migration tracks of U251 and U87 cells, respectively. Upper panels in a and b tracks of 10 randomly chosen cells; center panels images of migrating cells, and lower panels values of migration rate and mean distance based on tracks shown in upper panels . Values are means ± SD. ***Statistical relevance p < 0.001

Article Snippet: Human glioblastoma U251 MG (U251) and U87 MG (U87) cell lines were purchased from CLS Cell Lines Service (Germany) and monitored for correct genetic profile via microsatellite analyses according to a protocol described earlier (Peickert et al. ).

Techniques: Migration

Journal: Amino Acids

Article Title: Arginine deprivation affects glioblastoma cell adhesion, invasiveness and actin cytoskeleton organization by impairment of β-actin arginylation

doi: 10.1007/s00726-014-1857-1

Figure Lengend Snippet: Arginine deprivation impairs cell migration and invasiveness. Transwell filters not covered ( a ), and covered with Matrigel ( b , c) were used for analyses. Upper and lower panels in a and b images of U251 and U87 stained cells, respectively, taken on the filter trans side. c Images of LN-229 cells, analyzed as in b . Analyses were performed for three independent experiments run in duplicates. d Images of GFP-expressing U251 cells found within the E13 organotypic brain slice. The images represent the confocal 12.3-μm z -section of the planar center of brain slices. Right panel the quantification of GFP-expressing U251 cells within the confocal center of the slice per view area. The quantitative data in a – d are presented as % of control. Values are means ± SD. ***Statistical relevance p < 0.001

Article Snippet: Human glioblastoma U251 MG (U251) and U87 MG (U87) cell lines were purchased from CLS Cell Lines Service (Germany) and monitored for correct genetic profile via microsatellite analyses according to a protocol described earlier (Peickert et al. ).

Techniques: Migration, Staining, Expressing, Slice Preparation

Journal: Biochemistry and Biophysics Reports

Article Title: NF-κB signaling pathway mediates the anti-tumor effect of Ginsenoside Rg1 in glioblastoma

doi: 10.1016/j.bbrep.2026.102548

Figure Lengend Snippet: Evaluation of GS Rg1 Effect on the Viability and Proliferation of Human Glioblastoma U87-MG and U251-MG Cells using MTT and BrdU assays, Respectively. U87-MG(A and B) and U251-MG (C and D) cells were treated with 0, 5, 10 and 20 μM of GS Rg1 for 48 h, respectively. The viabilities were determined as percent compared to the controls. GS Rg1 inhibits proliferative properties of U87-MG and U251-MG cells in a concentration-dependent manner. The results are presented as mean ± SD from three independent experiments. (∗: p < 0.05, ∗∗: p < 0.01,and ∗∗∗: p < 0.001).

Article Snippet: The human glioma cell line U87-MG (U87 Malignant Glioma, #CL-0238; Wuhan, China) and U251-MG(U251 Malignant Glioma, #CL-0237; Wuhan, China) were obtained from Procell and cultured in Dulbecco's Modified Eagle's Medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Zhejiang Tianhang Biotechnology, Huzhou, China) and 1% antibiotics (Gibco), at 37 °C in a 5% CO 2 atmosphere.

Techniques: Concentration Assay

Journal: Biochemistry and Biophysics Reports

Article Title: NF-κB signaling pathway mediates the anti-tumor effect of Ginsenoside Rg1 in glioblastoma

doi: 10.1016/j.bbrep.2026.102548

Figure Lengend Snippet: Inhibition of MMP-2 and MMP-9 mRNA expression and activity of Extracellular Cathepsin B by GS Rg1 in U87-MG and U251-MG Cells. MMP2 (A and D) and MMP9 (B and E) expression levels were evaluated by reverse transcription-quantitative PCR. GS Rg1 at concentrations 5, 10 and 20 μM inhibited mRNA expression of MMP-2 and MMP-9, respectively. (C and F) Evaluation of GS Rg1 effect on proteolytic activity of extracellular Cathepsin B secreted from U87-MG and U251-MG cell line. GS Rg1 at concentrations 5, 10 and 20 μM inhibited activity of cathepsin, respectively. The values are given as mean ± SD compared with the control (∗: p < 0.05, ∗∗: p < 0.01, ∗∗∗: p < 0.001 and ∗∗∗∗: p < 0.0001).

Article Snippet: The human glioma cell line U87-MG (U87 Malignant Glioma, #CL-0238; Wuhan, China) and U251-MG(U251 Malignant Glioma, #CL-0237; Wuhan, China) were obtained from Procell and cultured in Dulbecco's Modified Eagle's Medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Zhejiang Tianhang Biotechnology, Huzhou, China) and 1% antibiotics (Gibco), at 37 °C in a 5% CO 2 atmosphere.

Techniques: Inhibition, Expressing, Activity Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Control

Journal: Biochemistry and Biophysics Reports

Article Title: NF-κB signaling pathway mediates the anti-tumor effect of Ginsenoside Rg1 in glioblastoma

doi: 10.1016/j.bbrep.2026.102548

Figure Lengend Snippet: Stimulation of Caspase-3 and Caspase-9 activity by GS Rg1 in U87-MG and U251-MG cells. (A and C) GS Rg1 increased activity of Caspase-3 in a concentration-dependent manner. (B and D) GS Rg1 increased activity of Caspase-9 in a concentration-dependent manner. The values are given as mean ± SD compared with the control (∗: p < 0.05, ∗∗: p < 0.01, and ∗∗∗: p < 0.001).

Article Snippet: The human glioma cell line U87-MG (U87 Malignant Glioma, #CL-0238; Wuhan, China) and U251-MG(U251 Malignant Glioma, #CL-0237; Wuhan, China) were obtained from Procell and cultured in Dulbecco's Modified Eagle's Medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Zhejiang Tianhang Biotechnology, Huzhou, China) and 1% antibiotics (Gibco), at 37 °C in a 5% CO 2 atmosphere.

Techniques: Activity Assay, Concentration Assay, Control

Journal: Biochemistry and Biophysics Reports

Article Title: NF-κB signaling pathway mediates the anti-tumor effect of Ginsenoside Rg1 in glioblastoma

doi: 10.1016/j.bbrep.2026.102548

Figure Lengend Snippet: Effect of GS Rg1 on transcriptional expression of proteins involved in Survival of Glioblastoma Cells. Transcriptional expression of IKK2(A and G), survivin (B and H), c-Myc (C and I), hTERT (D and J), NEMO (NF-κB regulators) (E and K), STAT3(F and L) in U87-MG and U251-MG cell line, after treatment with GS Rg1 for 48 h. Data are shown as fold change in relative expression compared with HPRT1 on the basis of comparative Ct (2-ΔΔCt) method. Values are shown as mean ± SD. Statistically different values of ∗: p < 0.05, ∗∗: p < 0.01, ∗∗∗: p < 0.001 and ∗∗∗∗: p < 0.0001 are determined compared with the control.

Article Snippet: The human glioma cell line U87-MG (U87 Malignant Glioma, #CL-0238; Wuhan, China) and U251-MG(U251 Malignant Glioma, #CL-0237; Wuhan, China) were obtained from Procell and cultured in Dulbecco's Modified Eagle's Medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Zhejiang Tianhang Biotechnology, Huzhou, China) and 1% antibiotics (Gibco), at 37 °C in a 5% CO 2 atmosphere.

Techniques: Expressing, Control

Journal: Biochemistry and Biophysics Reports

Article Title: NF-κB signaling pathway mediates the anti-tumor effect of Ginsenoside Rg1 in glioblastoma

doi: 10.1016/j.bbrep.2026.102548

Figure Lengend Snippet: Effect of GS Rg1 on transcriptional expression of proteins involved in apoptosis of Glioblastoma Cells. Transcriptional expression of Bax (A and I), Caspase-3 (B and J), Caspase-9 (C and K), Bcl-2 (D and L), Bax/Bcl-2 (E and M), Blf-1(F and N), Bcl-xl (G and O) and p21(H and P) in U87-MG and U251-MG cell line, after treatment with GS Rg1 for 48 h. Data are shown as fold change in relative expression compared with HPRT1 on the basis of comparative Ct (2-ΔΔCt) method. Values are shown as mean ± SD. Statistically different values of ∗: p < 0.05, ∗∗: p < 0.01, ∗∗∗: p < 0.001 and ∗∗∗∗: p < 0.0001 are determined compared with the control.

Article Snippet: The human glioma cell line U87-MG (U87 Malignant Glioma, #CL-0238; Wuhan, China) and U251-MG(U251 Malignant Glioma, #CL-0237; Wuhan, China) were obtained from Procell and cultured in Dulbecco's Modified Eagle's Medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Zhejiang Tianhang Biotechnology, Huzhou, China) and 1% antibiotics (Gibco), at 37 °C in a 5% CO 2 atmosphere.

Techniques: Expressing, Control

Journal: Biochemistry and Biophysics Reports

Article Title: NF-κB signaling pathway mediates the anti-tumor effect of Ginsenoside Rg1 in glioblastoma

doi: 10.1016/j.bbrep.2026.102548

Figure Lengend Snippet: Effect of GS Rg1 on transcriptional expression of proteins involved in invasion and media pH change of Glioblastoma Cells. Transcriptional expression of MMP14 (A and G), Cathepsin B (B and H), uPA (C and I), uPAR (D and J), CA9 (E and K) and NHE1(F and L) in U87-MG and U251-MG cell line, after treatment with GS Rg1 for 48 h. Data are shown as fold change in relative expression compared with HPRT1 on the basis of comparative Ct (2-ΔΔCt) method. Values are shown as mean ± SD. Statistically different values of ∗: p < 0.05, ∗∗: p < 0.01, ∗∗∗: p < 0.001 and ∗∗∗∗: p < 0.0001 are determined compared with the control.

Article Snippet: The human glioma cell line U87-MG (U87 Malignant Glioma, #CL-0238; Wuhan, China) and U251-MG(U251 Malignant Glioma, #CL-0237; Wuhan, China) were obtained from Procell and cultured in Dulbecco's Modified Eagle's Medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Zhejiang Tianhang Biotechnology, Huzhou, China) and 1% antibiotics (Gibco), at 37 °C in a 5% CO 2 atmosphere.

Techniques: Expressing, Control