Journal: Journal of visualized experiments : JoVE

Article Title: Cellular Membrane Affinity Chromatography Columns to Identify Specialized Plant Metabolites Interacting with Immobilized Tropomyosin Kinase Receptor B

doi: 10.3791/63118

Figure Lengend Snippet: Confocal microscopy images showing the immobilization of cell membrane fragments with functional TrkB receptors on IAM particles. (A) Cell membrane fragments from SH-SY5Y TrkB-NULL cell line immobilized on IAM particles after incubation with BDNF, primary antibody, and fluorophore-labeled secondary antibody. (B) Cell membrane fragments from SH-SY5Y Neuroblastoma cell lines expressing TrkB immobilized on IAM particles after incubation with primary antibody and fluorophore-labeled secondary antibody without BDNF. (C) Cell membrane fragments from SH-SY5Y neuroblastoma cell lines expressing TrkB immobilized on IAM particles after incubation with BDNF, primary antibody, and fluorophore-labeled secondary antibody.

Article Snippet: Cell culture of SH-SY5Y neuroblastoma cells (TrkB and TrkB-NULL (parental) cell lines) NOTE: Cell lines (SH-SY5Y Cell Line (TrkB, BR6) and SH-SY5Y Parental Cell Line (TrkB NULL)) 49 , 50 were purchased from Kerafast.

Techniques: Confocal Microscopy, Membrane, Functional Assay, Incubation, Labeling, Expressing

Journal: bioRxiv

Article Title: High Frequency MHz-Order Nanovibration Enables Cell Membrane Remodelling and Lipid Microdomain Manipulation

doi: 10.1101/2024.09.24.614713

Figure Lengend Snippet: (preceding page) : SRBW induced membrane lipid and membrane-associated pro- tein distribution . a Images (inset) and distribution of their GP values ( - 1 to +1; as represented by the colour bar) for untreated (control) and SRBW-irradiated (2.5 W for 5 mins) C-Laurdan-labelled SH-SY5Y neuroblastoma cells fixed at 0 and mins post-exposure incubation. The his- tograms for the GP values were fitted with Gaussian distributions, with the solid green curves representing the average between low (dashed blue curves) and high (dashed red curves) distri- butions. Scale bars denote a length of 30 µm. b High and c low GP coverage of both untreated and SRBW irradiated cells, obtained by resolving the GP histograms in both cases with the two best Gaussian fits. d Lipid raft and tau protein organisation in response to SRBW (2.5 W for 5 mins) at different post-exposure incubation times, and, e in the presence of BAPTA-AM and methyl-p’-cyclodextrin (Mp’CD); the control comprised cells unexposed to SRBW stimulation but incubated with media containing the same concentration in DMSO. Images were acquired at 100 ⇥ and 60 ⇥ magnification, respectively, and the scale bars denote 10 µm lengths. Cell nuclei were stained using NucBlue™ Live ReadyProbes™ (blue), tau proteins with anti-tau antibody and Alexa Fluor 488 secondary antibody (green), and, GM1 with Alexa Fluor™ 647 conjugate (red). The column on the right comprises a merger of these two channels. The data are represented in terms of its mean ± the standard error over triplicate runs, and the asterisks ⇤ , ⇤⇤ and ⇤⇤⇤⇤ indicate statistically significant differences with p < 0.05, p < 0.01 and p < 0.0001, respectively.

Article Snippet: Ltd. (Mount Waver- ley, VIC, Australia), whereas the SH-SY5Y neuroblastoma cell line was obtained through Dr Shwathy Ramesan at The Florey Institute of Neuroscience and Mental Health (Parkville, VIC, Australia).

Techniques: Membrane, Control, Irradiation, Incubation, Concentration Assay, Staining