memglow 488  (Cytoskeleton Inc)

Journal: medRxiv

Article Title: Detection and characterization of single SARS-CoV-2 viral particles by flow virometry

doi: 10.64898/2026.04.28.26351941

Figure Lengend Snippet: (A) Schematic representation of staining procedure for virus-containing supernatant. Supernatant from infected cells is harvested and centrifugated to remove cellular debris. Staining reagents are added to the sample, fixed with paraformaldehyde (PFA) and diluted in DPBS before being analyzed by flow virometry (FVM). (B) Graphical summary of different labelling techniques for targeting the SARS-CoV-2 spike protein via antibodies, the viral RNA genome via the nucleic acid intercalating dye Syto24 and the viral lipid envelope via the lipid intercalating dye MemGlow-488. (C) Dot plots and MFI of D614G stained with monoclonal antibody Tixagevimab (1 µg/mL; anti-RBD) conjugated to DyLight488 (DL488), with Syto24 (20 µM), with MemGlow-488 (200 nM), respectively.

Article Snippet: For staining of SARS-CoV-2 particles, cell culture supernatants were incubated with Tixagevimab conjugated to DyLight-488 (1 μg/mL) or Syto24 (20 μM; Thermo Fisher S7559) or MemGlow-488 (200 nM; Cytoskeleton #MG01) for 30 minutes at room temperature (RT).

Techniques: Staining, Virus, Infection

Journal: Redox Biology

Article Title: A clickable CoQ imaging probe reveals that cellular uptake and lysosomal trafficking depend on CD36 and NPC1

doi: 10.1016/j.redox.2025.103936

Figure Lengend Snippet: NPC1, CD36, LDLR involvement in CoQ uptake a ) Relative expression (log10-transformed RPKM value) of genes in murine brown adipocytes treated with 4CBA (24 h). b ) Relative expression (RPKM) of selected transcription factors in murine brown adipocytes treated with 4CBA or vehicle (CTL), based on RNA-seq analysis (n = 3 independent RNA pools per treatment). c ) Volcano plot obtained from DESeq2 analysis of vehicle control– and 4CBA-treated murine brown adipocyte RNA pools (n = 3 independent experiments per treatment). The Wald test was used for statistical analysis. d ) Strategy to verify NPC1, CD36, LDLR involvement in CoQ internalization by HPLC approach with CoQ 10 supplementation and imaging using CoQ Azide-SiR-DBCO SPAAC reaction. e ) Gene expression of NPC1, f ) CD36 and g ) LDLR siRNA-treated cells, n = 6. h-j ) CoQ 10 levels of murine brown adipocytes treated with vehicle or CoQ 10 ND (5 μM) for 24 h after transfection with siRNA, n = 6. Cells were treated on day 7, and harvested on day 8 for HPLC analysis. Data are presented as mean ± SEM. Results were compared using an unpaired two-tailed Student's t-test. Statistical significance is indicated as ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 and ∗∗∗∗P < 0.0001 compared to controls. Data represent biological replicates. For the colocalization analysis, BAT cells were treated with siRNAs as outlined in d . Cells were treated with 50 μM CoQ AzideND for 24 h at day 7 and analyzed the following day. SPAAC reaction and staining was performed as described in . To visualize the plasma membrane, the fluorescent probe MemGlow 488 (Cytoskeleton) was used at a final concentration of 1 μM after the SPAAC reaction. l ) First row, from left to right, representative confocal images of cells costained with CoQ Azide, Hoechst, and LysoTracker Green, along with m ) the corresponding colocalization analysis of CoQ Azide signal with lysosomes. n ) Costaining of CoQ Azide, Hoechst, and MitoTracker Green in the second row, and o ) respective colocalization of CoQ Azide signal with MitoTracker Green. p ) Third row, representative confocal images of cell costained with CoQ Azide, Hoechst and MEMGlow and q ) respective colocalization analysis of CoQ Azide signal with plasma membrane. Scale bar, 20 μm. Colocalization analysis was performed using JaCoP plugin on ImageJ. Data represent two independent experiments. A minimum of five images were analyzed for each condition. Results were compared using a one-way ANOVA test. Statistical significance is indicated as ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001.

Article Snippet: To visualize the plasma membrane, the fluorescent probe MemGlow 488 (Cytoskeleton) was used at a final concentration of 1 μM after the SPAAC reaction. l ) First row, from left to right, representative confocal images of cells costained with CoQ Azide, Hoechst, and LysoTracker Green, along with m ) the corresponding colocalization analysis of CoQ Azide signal with lysosomes. n ) Costaining of CoQ Azide, Hoechst, and MitoTracker Green in the second row, and o ) respective colocalization of CoQ Azide signal with MitoTracker Green. p ) Third row, representative confocal images of cell costained with CoQ Azide, Hoechst and MEMGlow and q ) respective colocalization analysis of CoQ Azide signal with plasma membrane.

Techniques: Expressing, Transformation Assay, RNA Sequencing, Control, Imaging, Gene Expression, Transfection, Two Tailed Test, Staining, Clinical Proteomics, Membrane, Concentration Assay