Journal: Cell reports

Article Title: mRNA decay pre-complex assembly drives timely cell-state transitions during differentiation

doi: 10.1016/j.celrep.2024.115138

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse cMyc (clone 9E10) (IF: 1:200) , MilliporeSigma , Cat#M4439; RRID: AB_439694.

Techniques: Recombinant, Staining, Software

Journal: Genome Biology and Evolution

Article Title: Evolutionary and Functional Analysis of Monoamine Oxidase F: A Novel Member of the Monoamine Oxidase Gene Family

doi: 10.1093/gbe/evae280

Figure Lengend Snippet: Expression of Indo-Pacific tarpon ( Megalops cyprinoides ) MAO A/B and MAO F in human cells. a and b) HeLa cells grown in glass coverslips were transfected to express full-length MAO A/B a) or MAO F b) tagged with three copies of the c-Myc epitope at the N-terminus (3myc-MAO A/B and 3myc-MAO F, respectively), followed by incubation with the mitochondrial probe MitoTracker Orange (red channel). Cells were fixed, permeabilized, and labeled with mouse monoclonal antibody against the c-Myc epitope, followed by incubation with Alexa-Fluor-488-conjugated donkey anti-mouse IgG (green channels), and nuclei were stained with the DNA probe DAPI. Stained cells were examined by fluorescence microscopy. In the merge channels, yellow indicates the colocalization of each MAO protein with MitoTracker at mitochondria, and red depicts mitochondria in nontransfected cells. Bar, 10 μm. c) The indicated cells were left untreated (Non-transfected; lanes 1 and 5), transfected with empty pCMV-3Tag-2A vector (lanes 2 and 6), or transfected with pCMV-3Tag-2A vector encoding either 3myc-MAO A/B (lanes 3 and 7) or 3myc-MAO F (lanes 4 and 8). Detergent-soluble extracts were prepared, and samples were processed by SDS-PAGE and immunoblot analysis with antibodies against the c-Myc epitope or antibody against β-ACTIN used as the loading control. The position of molecular mass markers is indicated on the left.

Article Snippet: The primary antibodies used were mouse monoclonal anti-Myc epitope (Covance, clone 9E10; 1:1,000 dilution) and mouse monoclonal anti-β-ACTIN used as an internal loading control (ThermoFisher, clone BA3R; 1:5,000 dilution).

Techniques: Expressing, Transfection, Incubation, Labeling, Staining, Fluorescence, Microscopy, Plasmid Preparation, SDS Page, Western Blot, Control

Journal: Genome Biology and Evolution

Article Title: Evolutionary and Functional Analysis of Monoamine Oxidase F: A Novel Member of the Monoamine Oxidase Gene Family

doi: 10.1093/gbe/evae280

Figure Lengend Snippet: Expression of Indo-Pacific tarpon ( Megalops cyprinoides ) MAO A/B and MAO F in human cells. a and b) HeLa cells grown in glass coverslips were transfected to express full-length MAO A/B a) or MAO F b) tagged with three copies of the c-Myc epitope at the N-terminus (3myc-MAO A/B and 3myc-MAO F, respectively), followed by incubation with the mitochondrial probe MitoTracker Orange (red channel). Cells were fixed, permeabilized, and labeled with mouse monoclonal antibody against the c-Myc epitope, followed by incubation with Alexa-Fluor-488-conjugated donkey anti-mouse IgG (green channels), and nuclei were stained with the DNA probe DAPI. Stained cells were examined by fluorescence microscopy. In the merge channels, yellow indicates the colocalization of each MAO protein with MitoTracker at mitochondria, and red depicts mitochondria in nontransfected cells. Bar, 10 μm. c) The indicated cells were left untreated (Non-transfected; lanes 1 and 5), transfected with empty pCMV-3Tag-2A vector (lanes 2 and 6), or transfected with pCMV-3Tag-2A vector encoding either 3myc-MAO A/B (lanes 3 and 7) or 3myc-MAO F (lanes 4 and 8). Detergent-soluble extracts were prepared, and samples were processed by SDS-PAGE and immunoblot analysis with antibodies against the c-Myc epitope or antibody against β-ACTIN used as the loading control. The position of molecular mass markers is indicated on the left.

Article Snippet: Cells were incubated with a monoclonal antibody to the c-Myc epitope (1:200 dilution; clone 9E10; Covance, Princeton, NJ, USA) for 30 min at 37 °C in a humidified chamber.

Techniques: Expressing, Transfection, Incubation, Labeling, Staining, Fluorescence, Microscopy, Plasmid Preparation, SDS Page, Western Blot, Control