human u87 glioblastoma  (ATCC)

Journal: Journal of Inflammation Research

Article Title: Synthetic 1,3,6-Tri-O-Galloyl-α-D-Glucose Mimics the Hippo Pathway Inhibitor VT107 in Suppressing Concanavalin A-Induced Inflammation in Human Glioblastoma Cells

doi: 10.2147/JIR.S565721

Figure Lengend Snippet: Hippo pathway inhibitors alter the Concanavalin A-induced phosphorylation of ERK and IκB in human U87 glioblastoma cells . Serum-starved human U87 glioblastoma cells were treated with 30 µg/mL Concanavalin A (ConA) for 5 min in the absence or presence of 3 µM IAG933 (IAG), GNE7883 (GNE), or VT107 (VT). ( A ) Protein cell lysates were harvested and processed for immunoblotting as described in the Methods section, to detect the phosphorylation status of ERK and of IκB. ( B ) Densitometric analysis was performed and phosphorylation status expressed as the extent of phospho-ERK/ERK or phosphor-IκB/IκB ratios over untreated control cells. Data points are the means ± SEM of 3 independent experiments.

Article Snippet: Human U87 glioblastoma (GBM)-derived cells were sourced from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Phospho-proteomics, Western Blot, Control

Journal: Journal of Inflammation Research

Article Title: Synthetic 1,3,6-Tri-O-Galloyl-α-D-Glucose Mimics the Hippo Pathway Inhibitor VT107 in Suppressing Concanavalin A-Induced Inflammation in Human Glioblastoma Cells

doi: 10.2147/JIR.S565721

Figure Lengend Snippet: Time-dependent inhibition of Concanavalin A-induced phosphorylation of ERK and IκB by αTGG in human U87 glioblastoma cells . Serum-starved human U87 glioblastoma cells were treated with 30 µg/mL Concanavalin A (ConA) for the indicated time in the absence or presence of 30 µM αTGG. ( A ) Protein cell lysates were harvested and processed for immunoblotting as described in the Methods section, to detect the phosphorylation status of ERK and of IκB. ( B ) Densitometric analysis was performed and phosphorylation status expressed as the maximal value of phospho-ERK/ERK or phosphor-IκB/IκB ratios for control (grey shades) vs αTGG (black shades). Western blot and densitometric analysis are representative of 3 independent experiments.

Article Snippet: Human U87 glioblastoma (GBM)-derived cells were sourced from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Inhibition, Phospho-proteomics, Western Blot, Control

Journal: Journal of Inflammation Research

Article Title: Synthetic 1,3,6-Tri-O-Galloyl-α-D-Glucose Mimics the Hippo Pathway Inhibitor VT107 in Suppressing Concanavalin A-Induced Inflammation in Human Glioblastoma Cells

doi: 10.2147/JIR.S565721

Figure Lengend Snippet: Dose-dependent inhibition of Concanavalin A-induced phosphorylation of ERK and IκB by αTGG in human U87 glioblastoma cells . Serum-starved human U87 glioblastoma cells were treated with the indicated concentrations of Concanavalin A (ConA) for 5 min in the absence or presence of 30 µM αTGG. ( A ) Protein cell lysates were harvested and processed for immunoblotting as described in the Methods section, to detect the phosphorylation status of ERK and of IκB. ( B ) A representative densitometric analysis was performed and phosphorylation status expressed as the maximal value of phospho-ERK/ERK or phosphor-IκB/IκB ratios for control (closed circles) vs αTGG (open circles). Western blot and densitometric analysis are representative of 3 independent experiments.

Article Snippet: Human U87 glioblastoma (GBM)-derived cells were sourced from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Inhibition, Phospho-proteomics, Western Blot, Control

Journal: Journal of Inflammation Research

Article Title: Synthetic 1,3,6-Tri-O-Galloyl-α-D-Glucose Mimics the Hippo Pathway Inhibitor VT107 in Suppressing Concanavalin A-Induced Inflammation in Human Glioblastoma Cells

doi: 10.2147/JIR.S565721

Figure Lengend Snippet: Dose-dependent inhibition of Concanavalin A-induced Snail, COX2, and activation of proMMP-2 by αTGG in human U87 glioblastoma cells . Serum-starved human U87 glioblastoma cells were treated with the indicated concentrations of αTGG in the absence or presence of 30 µg/mL Concanavalin A (ConA) for 24 hours. ( A ) Protein cell lysates were harvested and processed for immunoblotting as described in the Methods section, to detect the expression levels of Snail and COX2. GAPDH served as a loading control. ( B ) A gelatin zymography was performed using the conditioned media isolated from the respective conditions as described in the Methods section to assess the effect of αTGG on the extent of ConA-induced proMMP-2 activation. ( C ) U87 glioblastoma cells were treated with the indicated concentrations of αTGG for 24 hours. Cell viability was evaluated for each concentration using the Trypan Blue exclusion assay and a TC20 Automated Cell Counter (Biorad). Data are representative of two independent experiments.

Article Snippet: Human U87 glioblastoma (GBM)-derived cells were sourced from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Inhibition, Activation Assay, Western Blot, Expressing, Control, Zymography, Isolation, Concentration Assay, Trypan Blue Exclusion Assay

Journal: Journal of Inflammation Research

Article Title: Synthetic 1,3,6-Tri-O-Galloyl-α-D-Glucose Mimics the Hippo Pathway Inhibitor VT107 in Suppressing Concanavalin A-Induced Inflammation in Human Glioblastoma Cells

doi: 10.2147/JIR.S565721

Figure Lengend Snippet: Differential YAP/TEAD control of Concanavalin A-mediated regulation of Hippo pathway downstream effectors . Serum-starved human U87 glioblastoma cells were treated with the indicated concentrations of ConA for 24 hours, total RNA was extracted and RT-qPCR performed as described in the Methods section. ( A ) Gene expression of inflammatory ( COX2, IL6 ) and downstream Hippo pathway effectors ( AXL, CRGF, CYR61 , and NF2 ). ( B ) Transient gene silencing of YAP1 and TEAD was performed and extent and specificity of gene repression compared to PPIA . ( C ) Impact of YAP1 and TEAD silencing on ConA-mediated effects on downstream Hippo pathway effectors. Data points are the means ± SEM of triplicates from independent experiments.

Article Snippet: Human U87 glioblastoma (GBM)-derived cells were sourced from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Control, Quantitative RT-PCR, Gene Expression

Journal: Journal of Inflammation Research

Article Title: Synthetic 1,3,6-Tri-O-Galloyl-α-D-Glucose Mimics the Hippo Pathway Inhibitor VT107 in Suppressing Concanavalin A-Induced Inflammation in Human Glioblastoma Cells

doi: 10.2147/JIR.S565721

Figure Lengend Snippet: The Hippo pathway inhibitor VT107 alters dose-dependently the Concanavalin A-induced phosphorylation of ERK and IκB in human U87 glioblastoma cells . Serum-starved human U87 glioblastoma cells were treated with 30 µg/mL Concanavalin A (ConA) for 5 min in the absence or presence of the indicated concentrations of VT107 (VT). ( A ) Protein cell lysates were harvested and processed for immunoblotting as described in the Methods section, to detect the phosphorylation status of ERK and of IκB. ( B ) A representative densitometric analysis was performed and phosphorylation status expressed as the percent of phospho-ERK/ERK or phospho-IκB/IκB ratios of ConA-treated cells. Western blot and densitometric analysis are representative of 3 independent experiments. ( C ) Conditioned media was collected and gelatin zymography performed as described in the Methods section to monitor the impact of VT107 on proMMP-2 activation.

Article Snippet: Human U87 glioblastoma (GBM)-derived cells were sourced from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Phospho-proteomics, Western Blot, Zymography, Activation Assay

Journal: Journal of Inflammation Research

Article Title: Synthetic 1,3,6-Tri-O-Galloyl-α-D-Glucose Mimics the Hippo Pathway Inhibitor VT107 in Suppressing Concanavalin A-Induced Inflammation in Human Glioblastoma Cells

doi: 10.2147/JIR.S565721

Figure Lengend Snippet: The Hippo pathway inhibitor VT107 and aTGG share a common anti-inflammatory molecular signature in Concanavalin A-primed human U87 glioblastoma cells . ( A ) Serum-starved human U87 glioblastoma cells were treated for 24 hours with 30 µg/mL Concanavalin A (ConA), total RNA extracted and processed for a gene array screen of 141 inflammatory-associated genes as described in the Methods section by RT-qPCR. ( B ) Serum starved cells were similarly treated with ConA in the presence of 1 µM of either IAG933 (IAG), GNE7883 (GNE) or VT107 (VT) for 24 hours and extent of inhibition of the 10 most induced common genes assessed and correlated to that of 30 µM αTGG. ( C ) Protein-to-protein interaction network of the 10 commonly shared and downregulated genes between all the inhibitors tested in ConA-treated cells as determined with STRING.

Article Snippet: Human U87 glioblastoma (GBM)-derived cells were sourced from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Quantitative RT-PCR, Inhibition

Journal: Journal of Natural Products

Article Title: Paraisariamides: Cycloheptapeptide Toxins from Entomopathogenic Fungi ( Paraisaria spp.) That Inhibit Total Protein Synthesis

doi: 10.1021/acs.jnatprod.5c01111

Figure Lengend Snippet: Comparative biological activity of natural product Psds A–D ( 1 – 4 , 1 nM to 10 μM) from P. cascadensis relative to known elongation factor inhibitors, didemnin B (0.1 nM to 1 μM) and ternatin (0.1 nM to 1 μM). (A) Analysis of the secretory function of human U87-MG glioblastoma cells expressing the reporter Gaussia luciferase (GLuc). GLuc expression was assessed in the conditioned medium after 18 h in the presence of Psds 1 – 4 , didemnin B, ternatin, or vehicle (0.1% DMSO). (B) Viability of human HCT116 colon cancer cells and (C) wild-type human U87-MG glioblastoma cells exposed to pure compounds or 0.1% DMSO for 72 h. Cell viability was measured by quantification of ATP at the end point. Data points represent mean ± SE ( n = 3 wells per treatment), expressed as a % of vehicle-treated cells, from a comparison that was repeated at least three times.

Article Snippet: Human HeLa cervical cancer cells, human HCT116 colon cancer cells, and human U87-MG glioblastoma cells, were from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Activity Assay, Expressing, Luciferase, Comparison