c6 rat glioma cells american type culture collection  (ATCC)

Journal: Current cancer drug targets

Article Title: The Potential Targets and Mechanisms of a Carbazole and Pyrazole Containing Anticancer Compound

doi: 10.2174/1568009620666200115162343

Figure Lengend Snippet: VUGX01 induced caspase activities on cancer cell lines. All cells (1×104) were plated in multiple wells of 96-well plate. 24 hours after seeding, DMSO or indicated concentration of VUGX01 or Cetuximab were added to multiple wells of these cells. 24 hours later, the cell caspase activities were quantified by using Promega’s caspase-Glo kit. No drug treatment (DMSO alone) group was normalized as 1, data from HCT116, DLD1 and HEK293 treated with three concentrations (0.2, 1 and 5μM) were plotted in Panel A. In Panel B and C, VUGX01’s effect on Difi (B) or C6 (C) cells were compared to those from Cetuximab. All the data from the treatment group were normalized against their individual cell line’s non-treatment controls. Each data point was the average from 6 sample wells; standard deviations were shown as error bars. p-value of each data set is less than 0.05.

Article Snippet: Cell Culture Rat glioma C6 and human HEK293 cells were purchased from American Type Culture Collection (ATCC, Manassas, VA), and human colon cancer cells DLD1, Difi, and HCT116 were provided by one of our collaborators (Dr. Robert J Coffey, Vanderbilt University Medical Center, Nashville, TN).

Techniques: Concentration Assay

Journal: Current cancer drug targets

Article Title: The Potential Targets and Mechanisms of a Carbazole and Pyrazole Containing Anticancer Compound

doi: 10.2174/1568009620666200115162343

Figure Lengend Snippet: Western blots analysis. Whole-cell lysates were extracted from various cell lines (C6 in Panel A, HCT116, HEK293, DLD1, C6 in Panel B) treated with the indicated concentration of VUGX01 for 48 hours. 30μg of protein were separated by SDS-PAGE and blotted with antibodies against different antigens as labeled on the right. Tubulin was used as loading controls.

Article Snippet: Cell Culture Rat glioma C6 and human HEK293 cells were purchased from American Type Culture Collection (ATCC, Manassas, VA), and human colon cancer cells DLD1, Difi, and HCT116 were provided by one of our collaborators (Dr. Robert J Coffey, Vanderbilt University Medical Center, Nashville, TN).

Techniques: Western Blot, Concentration Assay, SDS Page, Labeling

Journal: Current cancer drug targets

Article Title: The Potential Targets and Mechanisms of a Carbazole and Pyrazole Containing Anticancer Compound

doi: 10.2174/1568009620666200115162343

Figure Lengend Snippet: GPCR Signaling Pathway Profiling. 82 selective genes in PathwayFinder RT2 Profiler PCR Array was screened by cDNA from DLD1 (control group) and VUGX01 treated group), their relative expression level was plotted against each other, genes corresponding to the spots outside triple lines (represented by gray light-colored spots) was considered to be significant in their expression difference (Panel A). Positive candidates were listed Panel B, they were further confirmed by another primer pairs by using cDNAs from rat C6 and three colon cancer cell lines. 9 genes marked with bold fonts were confirmed and showed significant change induced by VUGX01 treatment in all four cell lines.

Article Snippet: Cell Culture Rat glioma C6 and human HEK293 cells were purchased from American Type Culture Collection (ATCC, Manassas, VA), and human colon cancer cells DLD1, Difi, and HCT116 were provided by one of our collaborators (Dr. Robert J Coffey, Vanderbilt University Medical Center, Nashville, TN).

Techniques: Control, Expressing

Journal: Magnetic resonance in medicine

Article Title: Ischemia-Induced Changes of Intracellular Water Diffusion in Rat Glioma Cell Cultures

doi: 10.1002/mrm.21616

Figure Lengend Snippet: Preischemia 31P (a) and 1H (b) spectra of the HFBR C6 cell culture. Spectra were obtained at approximately 250 h of growth after inoculation. Labels in the 31P spectrum are: GPC, glycerophosphorylcholine; NTP, nucleoside triphosphates; Pi, inorganic phosphate; and PME, phosphomonoesters. The 1H spectrum was obtained before (black line) and after (gray line) the infusion of 5 mM Gd-DTPA into the media, which results in a splitting between intracellular (0 Hz) and extracellular (+185, +135 Hz) water components. The 1H spectrum obtained before Gd-DTPA was scaled down by a factor of eight for plotting.

Article Snippet: Cell Culture Rat glioma (C6) cells were obtained from American Type Culture collection and routinely cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS).

Techniques: Cell Culture

Journal: Magnetic resonance in medicine

Article Title: Ischemia-Induced Changes of Intracellular Water Diffusion in Rat Glioma Cell Cultures

doi: 10.1002/mrm.21616

Figure Lengend Snippet: 31P MRS spectra of the HFBR C6 culture as a function of time after the onset of ischemia. Labels refer to the time after ischemia in minutes. Peak assignments can be found in Fig. 1.

Article Snippet: Cell Culture Rat glioma (C6) cells were obtained from American Type Culture collection and routinely cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS).

Techniques:

Journal: Magnetic resonance in medicine

Article Title: Ischemia-Induced Changes of Intracellular Water Diffusion in Rat Glioma Cell Cultures

doi: 10.1002/mrm.21616

Figure Lengend Snippet: 1H MRS spectra of the HFBR C6 culture as a function of time after the onset of ischemia. Labels refer to the time after ischemia in minutes.

Article Snippet: Cell Culture Rat glioma (C6) cells were obtained from American Type Culture collection and routinely cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS).

Techniques:

Journal: Developmental cell

Article Title: The Dynein Adaptor RILP Controls Neuronal Autophagosome Biogenesis, Transport and Clearance

doi: 10.1016/j.devcel.2020.03.011

Figure Lengend Snippet: RILP was knocked down in DIV3 rat cortical neurons using RNAi and its effects on AP and LE behavior were monitored by time-lapse imaging of (C) RFP-LC3-positive APs, or (E) mCh-Rab7-positive LEs (n >20). Kymographs indicate the change in axonal AP and LE position over time in control (top) and RILP RNAi (bottom) neurons (scale bars: x = 5 μm; y = 30 sec). (A) Western blot showing depletion of RILP protein upon 72 hr RILP RNAi in rat C6 glioma cells, quantified in (B). Kymographs show processive retrograde transport of APs (C) and LEs (E) in control axons, which is inhibited upon RILP RNAi. (D) Quantitative analysis reveals specific RILP RNAi-mediated inhibition of retrograde AP and (F) LE transport. (G) Decreased number of APs upon RILP RNAi indicates a RILP role in AP formation. P-values: *=0.05, **=0.01 and ***=0.001.

Article Snippet: Cell Culture C6 rat glioma cells (ATCC # CCL-107) were cultured in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin (P/S), at 37°C with 5% C O2.

Techniques: Imaging, Control, Western Blot, Inhibition

Journal: Developmental cell

Article Title: The Dynein Adaptor RILP Controls Neuronal Autophagosome Biogenesis, Transport and Clearance

doi: 10.1016/j.devcel.2020.03.011

Figure Lengend Snippet: (A) Full-length GFP-RILP immunoprecipitated endogenous ATG5:12 from C6 glioma cell lysates, visualized by an anti-ATG5 antibody. (B) Full- length and N-terminal GST-RILP pulled down recombinant His- ATG5 in vitro. Live cell time-lapse imaging was used to monitor GFP-RILP behavior at mCh-ATG5-labeled isolation membranes in DIV7–8 cortical neurons. (C) Single frames for individual axons are shown at top for wt GFP-RILP and mCh-ATG5. Shown below are kymographs which indicate the positions of individual isolation membranes over time. Scale bars: x = 5μm; y= 30sec. RILP-positive isolation membranes were mostly immotile. (D) Immunostaining for dynein LIC1 subunit showed that ATG5-positive RILP structures were devoid of dynein (Top, (i)), while ATG5-negative RILP structures were enriched for dynein (bottom, (ii)) in soma as well as (E) neurites. (F) Percent relative localization of RILP with ATG5 and/or dynein. (G) Majority of RILP-positive isolation membranes are devoid of dynein. Scale bars: 5 μm. p-values: *=0.05, ***= 0.001.

Article Snippet: Cell Culture C6 rat glioma cells (ATCC # CCL-107) were cultured in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin (P/S), at 37°C with 5% C O2.

Techniques: Immunoprecipitation, Recombinant, In Vitro, Imaging, Labeling, Isolation, Immunostaining

Journal: Developmental cell

Article Title: The Dynein Adaptor RILP Controls Neuronal Autophagosome Biogenesis, Transport and Clearance

doi: 10.1016/j.devcel.2020.03.011

Figure Lengend Snippet: As a test for a physiological RILP role in autophagic turnover, we evaluated the distribution of Myc-tagged p62/sequestosome-1 (SQSTM1) with or without RILP RNAi in C6 cells. (A) p62 exhibited diffuse cytoplasmic distribution (top) in control cells. Upon RILP RNAi, p62 showed extensive aggregation (bottom), quantified in (B) (n>90). (C, D) GFP alone, wt RILP or LIR3-mutant RILP were transfected into C6 cells and endogenous p62 puncta were counted after 24 hours of expression (n=105 per condition). (C) Using immunostaining, comparable numbers of p62 puncta were detected in GFP and wt RILP expressing cells, but substantially higher numbers were observed in LIR3-mutant RILP expressing cells, quantified in (D). (E) Scatter plot shows relative increase in p62 aggregates upon RILP RNAi in C6 cells. (F-M) Further characterization of p62 aggregates shows that they are positive for endogenous LC3 (F) and ubiquitin (G), but mostly negative for Rab7 (H) and aggresome marker vimentin (I). Quantification of relative localization of p62 aggregates with LC3 (J), Ubiquitin (K), Rab7 (L) and Vimentin (M). P-value: ***=0.001.

Article Snippet: Cell Culture C6 rat glioma cells (ATCC # CCL-107) were cultured in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin (P/S), at 37°C with 5% C O2.

Techniques: Control, Mutagenesis, Transfection, Expressing, Immunostaining, Ubiquitin Proteomics, Marker

Journal: Developmental cell

Article Title: The Dynein Adaptor RILP Controls Neuronal Autophagosome Biogenesis, Transport and Clearance

doi: 10.1016/j.devcel.2020.03.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Cell Culture C6 rat glioma cells (ATCC # CCL-107) were cultured in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin (P/S), at 37°C with 5% C O2.

Techniques: Ubiquitin Proteomics, Recombinant, Protease Inhibitor, Mutagenesis, shRNA, Plasmid Preparation, Software