Journal: Fluids and Barriers of the CNS

Article Title: Induced pluripotent stem cell-derived cells model brain microvascular endothelial cell glucose metabolism

doi: 10.1186/s12987-022-00395-z

Figure Lengend Snippet: Western blot antibodies

Article Snippet: IDH1 , Cell Signaling Technology , 8137S.

Techniques: Western Blot

Journal: NPJ Precision Oncology

Article Title: Secondary IDH1 resistance mutations and oncogenic IDH2 mutations cause acquired resistance to ivosidenib in cholangiocarcinoma

doi: 10.1038/s41698-022-00304-5

Figure Lengend Snippet: a Immunoblot analysis of IDH1 expression in TF-1 cells infected with lentivirus expressing the indicated IDH1 variants and treated with vehicle-control (DMSO), 500 nM ivosidenib (IVO), or 500 nM LY3410738 (LY), as indicated. b , c Quantification of (R) -2HG ( b ) and fold-change in (R) -2HG ( c ) in the TF-1 cell lines shown in a , as indicated. Shown are mean values (±SD) of duplicate experiments. d , e Proliferation under cytokine-poor conditions ( d ) and fold-change in day 15 cell counts ( e ) of the TF-1 cells shown in ( a ) The mean (±SD) cell counts of three replicates is shown. f Fold-change in intracellular (R) -2HG in TF-1 cells expressing IDH1 R132H (left) or IDH1 R132H/D279N (right) treated overnight with the indicated concentrations of LY3410738. Shown are mean values (±SD) of duplicate experiments. In all cases, representative results from at least two independent experiments are shown.

Article Snippet: Primary antibodies used: rabbit monoclonal HA-tag (Cell Signaling Technology, #3724) at a 1:1000 dilution, rabbit polyclonal anti-IDH1 (Cell Signaling Technology, #3997) at a 1:500 dilution, and mouse monoclonal anti-vinculin (Sigma, V9131) at a 1:1000 dilution.

Techniques: Western Blot, Expressing, Infection

Journal: NPJ Precision Oncology

Article Title: Secondary IDH1 resistance mutations and oncogenic IDH2 mutations cause acquired resistance to ivosidenib in cholangiocarcinoma

doi: 10.1038/s41698-022-00304-5

Figure Lengend Snippet: a Predicted binding mode of ivosidenib (IVO, carbons in yellow) in the allosteric site of IDH1 R132C (carbons in blue). IVO is recognized by IDH1 R132C principally by donating a hydrogen bond to the IDH1 D279 side chain and accepting a hydrogen bond from the IDH1 A111 backbone. Further specific intermolecular interactions involve aromatic contacts with IDH1 W267, IDH1 W124, and IDH1 Y285. b IVO is predicted to have impaired binding to IDH1 R132C/D279N (carbons in red) because the chlorophenyl ring clashes with the mutated asparagine side chain (orange X). c Predicted binding mode of LY3410738 (LY, carbons in green) in the allosteric site of IDH1 R132C (carbons in blue). LY3410738 covalently binds to IDH1 C269 and displays many favorable electrostatic interactions with the protein: the protonated piperazine makes an ionic interaction with the IDH1 D279 side chain and the inhibitor additionally makes two hydrogen bonds with the IDH1 I128 backbone and accepts a hydrogen bond from the IDH1 L120 backbone. d LY3410738 is predicted to adopt a similar covalent binding mode to IDH1 R132C/D279N as it does to IDH1 R132C by keeping all of its numerous anchor points and by replacing the ionic interaction with aspartate with a hydrogen bond to the oxygen of the asparagine-mutated side chain at position 279.

Article Snippet: Primary antibodies used: rabbit monoclonal HA-tag (Cell Signaling Technology, #3724) at a 1:1000 dilution, rabbit polyclonal anti-IDH1 (Cell Signaling Technology, #3997) at a 1:500 dilution, and mouse monoclonal anti-vinculin (Sigma, V9131) at a 1:1000 dilution.

Techniques: Binding Assay

Journal: iScience

Article Title: Targeting HOTAIRM1 ameliorates glioblastoma by disrupting mitochondrial oxidative phosphorylation and serine metabolism

doi: 10.1016/j.isci.2022.104823

Figure Lengend Snippet:

Article Snippet: IDH1 Rabbit polyclonal antibody , Cell Signaling Technology , # 3997; RRID: AB_1904011.

Techniques: Labeling, Recombinant, Transfection, Protease Inhibitor, Plasmid Preparation, Isolation, Reporter Assay, Microarray, shRNA, Negative Control, Software

Journal: Frontiers in Oncology

Article Title: Cross-Talk Between Histone Methyltransferases and Demethylases Regulate REST Transcription During Neurogenesis

doi: 10.3389/fonc.2022.855167

Figure Lengend Snippet: Antibodies for ChIP.

Article Snippet: Acetyl H3K27 , 8137s , Cell Signaling Technology.

Techniques:

Journal: PLoS ONE

Article Title: Cancer-associated IDH mutations induce Glut1 expression and glucose metabolic disorders through a PI3K/Akt/mTORC1-Hif1α axis

doi: 10.1371/journal.pone.0257090

Figure Lengend Snippet: (A–C) Induction of Ptgs2 (A), Lamc2 (B), and Slc2a1 (C) in MEF cells stably expressing the IDH1 or IDH2 mutant. Expression was determined by real time-PCR. Expression of Gapdh was used as an internal control. The data represent mean ± SD of three independent experiments. * p<0.05, ** p<0.01, *** p<0.001. (D–F) Induction of Ptgs2 (D), Lamc2 (E), and Slc2a1 (F) in MEF cells treated with 300 μM of 2-HG for 24 h. ** p<0.01, *** p<0.001. (G) Elevated expression of Glut1 protein by the IDH1/2 mutants. Expression of β-actin served as an internal control. The data are representative of three independent experiments. (H) Elevated expression of Glut1 protein in response to the treatment with 300 μM of 2-HG. Cell lysates were harvested at 24 h or 48 h after treatment and subjected to Western blotting.

Article Snippet: After the blocking with 5% skim milk in TBS-T (Tris-buffered saline-Tween20) for 1 h, the membranes were incubated overnight with primary antibodies including anti-Flag (F3165, Sigma), anti-Glut1 (ab115730, Abcam, Cambridge, UK), anti-phospho-S6k (9205, Cell Signaling Technology, Danvers, MA), anti-total-S6k (2708, Cell Signaling Technology), anti-phospho-Akt (Ser473) (4060, Cell Signaling Technology), anti-phospho-Akt (Thr308) (13038, Cell Signaling Technology), anti-total-Akt (4681, Cell Signaling Technology), anti-Hif1α (14179, Cell Signaling Technology), anti-IDH1 (D2H1) (8137, Cell Signaling Technology), anti-IDH2 (D8E3B) (56439, Cell Signaling Technology), and anti-β-actin (A5441, Sigma).

Techniques: Stable Transfection, Expressing, Mutagenesis, Real-time Polymerase Chain Reaction, Western Blot

Journal: The Journal of Veterinary Medical Science

Article Title: Identification of regulated proteins by epigallocatechin gallate treatment in an ischemic cerebral cortex animal model: a proteomics approach

doi: 10.1292/jvms.21-0089

Figure Lengend Snippet: Sequence of the primers used for PCR amplification

Article Snippet: The list of primary antibodies is as follows: anti-isocitrate dehydrogenase (ICDH, #8137S; Cell Signaling Technology, Beverly, MA, USA), anti-dynamin-like protein 1 (DLP-1, 611112; BD Bioscience, Franklin Lakes, NJ, USA), anti-γ-enolase (sc-31859; Santa Cruz Biotechnology, Dallas, TX, USA), anti-chronophin/PDXP (Cell Signaling Technology, #4686), anti-60 kDa heat shock protein (HSP60, Cell Signaling Technology, #4870S), (diluted 1:1,000 with TBST), and anti-β-actin (Santa Cruz Biotechnology, sc-4778).

Techniques: Sequencing

Journal: The Journal of Veterinary Medical Science

Article Title: Identification of regulated proteins by epigallocatechin gallate treatment in an ischemic cerebral cortex animal model: a proteomics approach

doi: 10.1292/jvms.21-0089

Figure Lengend Snippet: List of identified proteins that were significantly differentially expressed in vehicle- and epigallocatechin gallate (EGCG)-treated animals with middle cerebral artery occlusion

Article Snippet: The list of primary antibodies is as follows: anti-isocitrate dehydrogenase (ICDH, #8137S; Cell Signaling Technology, Beverly, MA, USA), anti-dynamin-like protein 1 (DLP-1, 611112; BD Bioscience, Franklin Lakes, NJ, USA), anti-γ-enolase (sc-31859; Santa Cruz Biotechnology, Dallas, TX, USA), anti-chronophin/PDXP (Cell Signaling Technology, #4686), anti-60 kDa heat shock protein (HSP60, Cell Signaling Technology, #4870S), (diluted 1:1,000 with TBST), and anti-β-actin (Santa Cruz Biotechnology, sc-4778).

Techniques: Sequencing

Journal: The Journal of Veterinary Medical Science

Article Title: Identification of regulated proteins by epigallocatechin gallate treatment in an ischemic cerebral cortex animal model: a proteomics approach

doi: 10.1292/jvms.21-0089

Figure Lengend Snippet: Magnified protein spots of isocitrate dehydrogenase (ICDH), dynamin-like protein 1 (DLP-1), γ-enolase, pyridoxal-5′-phosphate phosphatase (PLPP), and 60 kDa heat shock protein (HSP60) in the cerebral cortex from vehicle + sham, epigallocatechin gallate (EGCG) + sham, vehicle + middle cerebral artery occlusion (MCAO), and EGCG + MCAO animals. Each square indicates the protein spots.

Article Snippet: The list of primary antibodies is as follows: anti-isocitrate dehydrogenase (ICDH, #8137S; Cell Signaling Technology, Beverly, MA, USA), anti-dynamin-like protein 1 (DLP-1, 611112; BD Bioscience, Franklin Lakes, NJ, USA), anti-γ-enolase (sc-31859; Santa Cruz Biotechnology, Dallas, TX, USA), anti-chronophin/PDXP (Cell Signaling Technology, #4686), anti-60 kDa heat shock protein (HSP60, Cell Signaling Technology, #4870S), (diluted 1:1,000 with TBST), and anti-β-actin (Santa Cruz Biotechnology, sc-4778).

Techniques:

Journal: The Journal of Veterinary Medical Science

Article Title: Identification of regulated proteins by epigallocatechin gallate treatment in an ischemic cerebral cortex animal model: a proteomics approach

doi: 10.1292/jvms.21-0089

Figure Lengend Snippet: Western blot analysis of isocitrate dehydrogenase (ICDH), dynamin-like protein 1 (DLP-1), γ-enolase, pyridoxal-5′-phosphate phosphatase (PLPP), and 60 kDa heat shock protein (HSP60) in the cerebral cortex from vehicle + sham, epigallocatechin gallate (EGCG) + sham, vehicle + middle cerebral artery occlusion (MCAO), and EGCG + MCAO animals. Densitometric analysis is represented as a ratio of β-actin intensity. Data ( n =5 per group) are represented as mean ± S.E.M. # P <0.05 vs. vehicle + sham animals, * P <0.05 vs. vehicle + MCAO animals.

Article Snippet: The list of primary antibodies is as follows: anti-isocitrate dehydrogenase (ICDH, #8137S; Cell Signaling Technology, Beverly, MA, USA), anti-dynamin-like protein 1 (DLP-1, 611112; BD Bioscience, Franklin Lakes, NJ, USA), anti-γ-enolase (sc-31859; Santa Cruz Biotechnology, Dallas, TX, USA), anti-chronophin/PDXP (Cell Signaling Technology, #4686), anti-60 kDa heat shock protein (HSP60, Cell Signaling Technology, #4870S), (diluted 1:1,000 with TBST), and anti-β-actin (Santa Cruz Biotechnology, sc-4778).

Techniques: Western Blot

Journal: The Journal of Veterinary Medical Science

Article Title: Identification of regulated proteins by epigallocatechin gallate treatment in an ischemic cerebral cortex animal model: a proteomics approach

doi: 10.1292/jvms.21-0089

Figure Lengend Snippet: Reverse transcription-PCR (RT-PCR) of isocitrate dehydrogenase (ICDH), dynamin-like protein 1 (DLP-1), γ-enolase, pyridoxal-5′-phosphate phosphatase (PLPP), and 60 kDa heat shock protein (HSP60) in the cerebral cortex from vehicle + sham, epigallocatechin gallate (EGCG) + sham, vehicle + middle cerebral artery occlusion (MCAO), and EGCG + MCAO animals. The band intensity of the RT-PCR product is expressed as a ratio of β-actin product intensity. Data ( n =5 per group) are shown as mean ± S.E.M. # P <0.05 vs. vehicle + sham animals, * P <0.05 vs. vehicle + MCAO animals.

Article Snippet: The list of primary antibodies is as follows: anti-isocitrate dehydrogenase (ICDH, #8137S; Cell Signaling Technology, Beverly, MA, USA), anti-dynamin-like protein 1 (DLP-1, 611112; BD Bioscience, Franklin Lakes, NJ, USA), anti-γ-enolase (sc-31859; Santa Cruz Biotechnology, Dallas, TX, USA), anti-chronophin/PDXP (Cell Signaling Technology, #4686), anti-60 kDa heat shock protein (HSP60, Cell Signaling Technology, #4870S), (diluted 1:1,000 with TBST), and anti-β-actin (Santa Cruz Biotechnology, sc-4778).

Techniques: Reverse Transcription Polymerase Chain Reaction