Journal: Biochemical and biophysical research communications

Article Title: Regulation of calcium release from the endoplasmic reticulum by the serine hydrolase ABHD2

doi: 10.1016/j.bbrc.2017.06.195

Figure Lengend Snippet: Proteins identified as targets of KT195

Article Snippet: Clones isolated by limiting dilution were screened for levels of ABHD2 protein on Western blots and mRNA by real-time PCR (probe Mm00502098_m1, ThermoFisher Scientific), as previously described [ 18 ].

Techniques:

Journal: Biochemical and biophysical research communications

Article Title: Regulation of calcium release from the endoplasmic reticulum by the serine hydrolase ABHD2

doi: 10.1016/j.bbrc.2017.06.195

Figure Lengend Snippet: Subcellular fractions (cytosol (C), ER, MAM, mitochondria (M)) and whole cell lysates (L) were prepared from IMLF and analyzed for protein markers by Western blot analysis using antibodies to ABHD2, fatty acid CoA ligase 4 (FACL4), oxidative phosphorylation complex V (OxPhos), MEK1/2 and GRP78.

Article Snippet: Clones isolated by limiting dilution were screened for levels of ABHD2 protein on Western blots and mRNA by real-time PCR (probe Mm00502098_m1, ThermoFisher Scientific), as previously described [ 18 ].

Techniques: Western Blot, Phospho-proteomics

Journal: Biochemical and biophysical research communications

Article Title: Regulation of calcium release from the endoplasmic reticulum by the serine hydrolase ABHD2

doi: 10.1016/j.bbrc.2017.06.195

Figure Lengend Snippet: IMLF were transfected with lentiviruses containing ABHD2 shRNA or control vector and stable clones generated. (A) LDH release was measured from ABHD2 shRNA-knockdown IMLF (triangles) and vector controls (circles) 30 min after stimulation with the indicated concentrations of A23187 (n=3, *P<0.05). A Western blot (inset) shows that ABHD2 shRNA-knockdown IMLF express less ABHD2 than vector controls. G-CEPIAer or CEPIAmt was expressed in vector control (solid line) and ABHD2 shRNA-knockdown IMLF (hatched line) to measure relative changes in ER (B) and mitochondrial (C) calcium, respectively. Calcium transients were measure after addition of A23187 (1 μg/ml) by live-cell fluorescent imaging. The data represent the average of three independent experiments from analysis of at least 5 cells/experiment.

Article Snippet: Clones isolated by limiting dilution were screened for levels of ABHD2 protein on Western blots and mRNA by real-time PCR (probe Mm00502098_m1, ThermoFisher Scientific), as previously described [ 18 ].

Techniques: Transfection, shRNA, Control, Plasmid Preparation, Clone Assay, Generated, Knockdown, Western Blot, Imaging