Journal: The Journal of Neuroscience

Article Title: Cholinergic Control of GnRH Neuron Physiology and Luteinizing Hormone Secretion in Male Mice: Involvement of ACh/GABA Cotransmission

doi: 10.1523/JNEUROSCI.1780-23.2024

Figure Lengend Snippet: An overview of the reagents used for immunohistochemistry

Article Snippet: At the same time, to detect Cy5 emission, a He–Ne laser (637 nm) and a DAPI/Cy5 (custom 239415; Semrock) dual emission filters were applied.

Techniques: Immuno-Electron Microscopy, Electron Microscopy

Journal:

Article Title: Inefficient degradation of truncated polyglutamine proteins by the proteasome

doi: 10.1038/sj.emboj.7600426

Figure Lengend Snippet: The proteasome is stably associated with aggregates of mutant Huntingtin and simple polyglutamine expansion proteins. (A) Colocalization of the proteasome with Htt-Q65 or Flag-Q81 aggregates. (Upper panel) HeLa cells were transfected with constructs encoding LMP2-GFP together with Htt-Q23, Htt-Q65, or Flag-Q81 as indicated. The Htt proteins and Flag-Q81 were detected with the HP-1 antibody and an antibody against the Flag epitope, respectively, followed by a TRITC-conjugated secondary antibody. Cells expressing LMP2-GFP (GFP, green) and Htt proteins or Flag-Q81 (TRITC, red) were visualized by fluorescence microscopy and phase contrast microscopy (Phase). Colocalization was illustrated by merging GFP and TRITC images (Merge). DNA was stained with DAPI (DAPI). (Lower panel) Localization of endogenous proteasome in the absence or presence of Flag-Q81 expression was detected using an anti-20S proteasome antibody followed by an FITC-conjugated secondary antibody (FITC, green). Scale bar represents 10 μm. (B) FRAP analysis of LMP2-GFP. Cells were imaged before photobleaching (Pre) of the defined area (white box) and at the indicated times after photobleaching. Note that the intensity of the images is scaled differently between the samples. Scale bar represents 5 μm. Quantitative FRAP analysis of soluble or aggregate-associated LMP2-GFP is shown in the graph in the right panel. The relative fluorescence intensity (RFI) was determined for each time point and is represented as the average±s.e.m. of 5–8 cells. (C) FLIP analysis of LMP2-GFP. Single scan images were obtained before (Pre) and at the indicated times between consecutive bleach pulses of the boxed area (white box). Scale bar represents 5 μm. The RFI was determined for each time point and is represented as the average±s.e.m. of five cells (right panel).

Article Snippet: FITC, Texas red, DAPI, and YFP filter sets (Chroma Technology Corp.) were used for visualization.

Techniques: Stable Transfection, Mutagenesis, Transfection, Construct, FLAG-tag, Expressing, Fluorescence, Microscopy, Staining