64169s  (Cell Signaling Technology Inc)

Journal: Frontiers in Cellular Neuroscience

Article Title: Investigations into photoreceptor energy metabolism during experimental retinal detachment

doi: 10.3389/fncel.2022.1036834

Figure Lengend Snippet: Primary antibodies used in the study.

Article Snippet: AGC1 , Cell Signaling Technology , cat# 64169 , Rabbit , Synthetic peptide corresponding to residues surrounding Arg309 of human AGC1 , 1:1,000.

Techniques: Purification, Isolation, Sequencing, Recombinant, Membrane

Journal: Frontiers in Cellular Neuroscience

Article Title: Investigations into photoreceptor energy metabolism during experimental retinal detachment

doi: 10.3389/fncel.2022.1036834

Figure Lengend Snippet: Effect of retinal detachment on expression of six mitochondrial enzymes within the same 3 days detached retina. (A) Cytochrome C oxidase (COX IV). (B) Succinate-CoA ligase (SUCLA2). (C) Aspartate-glutamate carrier 1 (AGC1). (D) Mitochondrial aspartate aminotransferase (mAST). (E) Mitochondrial superoxide dismutase (SOD2). (F) Mitochondrial creatine kinase (CK-MT1A). Scale bar: overviews = 150 μm; insets = 60 μm. INL, inner nuclear layer; SI, inner segments; ONL, outer nuclear layer.

Article Snippet: AGC1 , Cell Signaling Technology , cat# 64169 , Rabbit , Synthetic peptide corresponding to residues surrounding Arg309 of human AGC1 , 1:1,000.

Techniques: Expressing

Journal: Frontiers in cellular neuroscience

Article Title: Investigations into photoreceptor energy metabolism during experimental retinal detachment.

doi: 10.3389/fncel.2022.1036834

Figure Lengend Snippet: FIGURE 3 Effect of retinal detachment on expression of six mitochondrial enzymes within the same 3 days detached retina. (A) Cytochrome C oxidase (COX IV). (B) Succinate-CoA ligase (SUCLA2). (C) Aspartate-glutamate carrier 1 (AGC1). (D) Mitochondrial aspartate aminotransferase (mAST). (E) Mitochondrial superoxide dismutase (SOD2). (F) Mitochondrial creatine kinase (CK-MT1A). Scale bar: overviews = 150 µm; insets = 60 µm. INL, inner nuclear layer; SI, inner segments; ONL, outer nuclear layer.

Article Snippet: AGC1 Cell Signaling Technology cat# 64169 Rabbit Synthetic peptide corresponding to residues surrounding Arg309 of human AGC1 1:1,000 mAST Santa-Cruz cat# sc-271702 Mouse aa.

Techniques: Expressing

Journal: bioRxiv

Article Title: SLC25A39 is necessary for mitochondrial glutathione import in mammalian cells

doi: 10.1101/2021.09.15.460381

Figure Lengend Snippet: A. Relative abundance of GSH in whole cells (top) and mitochondria (bottom) immune isolated from HeLa cells treated with BSO (1 mM) for the indicated times compared to untreated controls. Bars represent mean ± s.d.; n = 3 biologically independent samples. Statistical significance was determined by one-way ANOVA followed by Bonferroni post-hoc analysis. B. Schematics of proteomics analysis of mitochondria isolated from indicated HeLa cells. C. Volcano plot showing relative fold change (log 2 ) in mitochondrial protein abundance vs P values (-log) from HeLa cells treated with BSO (1 mM) for 4 days compared to untreated (top). The dotted line represents P =0.01. Colors denote indicated protein families. Gene ontology analysis of proteins altered by BSO treatment (bottom). D. Immunoblot analysis of indicated proteins in whole-cell lysates and mitochondria isolated from HeLa cells treated with BSO (1 mM) for the indicated days. E. Immunoblot of indicated proteins in the HEK-293T (top) and K-562 (bottom) cells treated with BSO (1 mM;10 μM, respectively) for the indicated days. CS and GAPDH were used as loading controls. F. Immunofluorescence analysis of indicated proteins in HeLa cells treated with vehicle or BSO (1 mM) after 24 hours. Micrographs are representative images. Scale bar, 10 μm. G. Immunoblot of indicated proteins in the HeLa cells infected with the indicated sgRNAs. Cells were plated 6 days after transduction and treated for 4 days with GSHee (10 mM). SLC25A12 and β-ACTIN were used as loading controls. H. Immunoblot of indicated proteins in the HeLa cells treated with BSO (1 mM) and co-treated with either GSH (10 mM), GSHee (10 mM), Trolox (50 μM) or Ferrostatin-1 (5 μM) for 48 hours. SLC25A12 and CS were used as loading controls.

Article Snippet: Antibodies against beta-Actin (GTX109639), GAPDH (GTX627408) and PPAT (GTX102725) were obtained from GeneTex; lipoic acid antibody (437695) from EMD Millipore; SLC25A39 (14963-1-AP), ALAS1 (16200-1-AP), GCLC (12601-1-AP), HMOX1 (10701-1-AP), MRM1 (16392-1-AP), MRPL15 (18339-1-AP), MRPS7 (26828-1-AP), FDX1 (12592-1-AP), FECH (14466-1-AP) and LIAS (11577-1-AP) antibodies from Proteintech; anti-FLAG M2 (F1804) from Sigma; total OXPHOS antibody cocktail (ab110411) and SDHB (ab14714) antibody from Abcam; NFS1 (sc-365308) from Santa Cruz Biotechnology; SLC25A12 (64169S), calreticulin (12238P), RCAS1 (12290S), LAMP1 (9091P), CS (14309S), ACO2 (6571S), IRP2 (37135S), TFRC (13208S), FTH1 (3998S), cleaved PARP (9546S), SLC7A11 (12691S) and RPS6 (2217S) antibodies from CST; and ATP5A (43-9800) from ThermoFisher Scientific.

Techniques: Isolation, Quantitative Proteomics, Western Blot, Immunofluorescence, Infection, Transduction