prolong gold antifade mountant with dna stain dapi  (Thermo Fisher)

Journal: bioRxiv

Article Title: Multivalent Lipid MVL5 Micellar Nanoparticles Exhibit Dramatically Increased Loading of Paclitaxel with PEGylation Enhancing Human Cancer Cell Penetration Depth and Cytotoxicity

doi: 10.1101/2025.05.24.655953

Figure Lengend Snippet: M21 cells were treated with CLNPs of molar composition DOTAP/DOPC/PEG-lipid/TRITC-DHPE/Janelia-PTX equal to 50/46.5−X PEG /X PEG /0.5/3 mol% with either 0 ( a ) or 10 ( c ) mol% PEG-lipid and CLNPs with composition MVL5/DOPC/PEG-lipid/TRITC-DHPE/Janelia-PTX equal to 50/46.5−X PEG /X PEG /0.5/3 mol% with 0 ( b ) or 10 ( d ) mol% PEG-lipid. CLNP lipid (TRITC-DHPE) signal is shown in green, PTX (Janelia-PTX) signal is shown in red, DAPI was used to stain cell nuclei after fixation and is shown in blue. The main image at the top left panel of each image set shows an X-Y section at a selected Z from a Z-stack image series which intersects the body of the cell. The right projection shows the Y-Z side view of the slice along the vertical orange line in the main image. The bottom projection shows the X-Z side view of the slice along the horizontal orange line in the main image. Scale bars on the main image and projections are 20 mm and 2.5mm, respectively.

Article Snippet: ProLong Gold Antifade Mountant with DAPI DNA Stain was purchased from Fisher Scientific.

Techniques: Staining

Journal: bioRxiv

Article Title: Multivalent Lipid MVL5 Micellar Nanoparticles Exhibit Dramatically Increased Loading of Paclitaxel with PEGylation Enhancing Human Cancer Cell Penetration Depth and Cytotoxicity

doi: 10.1101/2025.05.24.655953

Figure Lengend Snippet: Images of M21 cells treated with CLNPs of molar composition MVL5/DOPC/PEG-lipid/TRITC- DHPE/Janelia-PTX equal to 50/46.5−X PEG /X PEG /0.5/3 mol % and DOTAP/DOPC/PEG-lipid/TRITC- DHPEE/Janelia-PTX equal to 50/46.5−X PEG /X PEG /0.5/3 mol % with either 0 or 10 mol % PEG-lipid are shown in ( a ). CLNP lipid (TRITC-DHPE) signal is shown in green, PTX (Janelia-PTX) signal is shown in red, DAPI was used to stain cell nuclei after fixation and is shown in blue. Scale bars are 50 μm.. The number of lipid puncta observed within the border of the cells treated with each formulation at this z-slice is quantified in ( b ) using the Cell Profiler particle (speckles) analysis. The distance of the identified lipid puncta from the cell border for cells treated with each formulation is quantified in ( c ). Statistical significance was determined using Dunn’s test in R and shown by asterisks (* for P<0.05, ** for P<0.01, *** for P<0.001, **** for P<0.0001). In ( b ), a box plot is shown where the ranked puncta per cell data for each formulation is binned into quartiles, the range of which are shown as vertical lines for the first and fourth quartiles, and the second and third quartiles are shown as a box, with the median of data shown as a horizontal line. Outliers were determined as points greater than 1.5 times the range of the second and third quartile away from the start of the fourth quartile and are shown as individual dots. In ( c ) a violin plot of the ranked distance of puncta to cell edge data, where a gaussian curve estimating the relative density of data points at each distance value along the y-axis is plotted, is overlayed on a box plot of the same data set.

Article Snippet: ProLong Gold Antifade Mountant with DAPI DNA Stain was purchased from Fisher Scientific.

Techniques: Staining, Formulation

Journal: Viruses

Article Title: The Virus Entry Pathway Determines Sensitivity to the Antiviral Peptide TAT-I24

doi: 10.3390/v17040458

Figure Lengend Snippet: Effect of peptides on dsRNA levels in infected Vero E6 cells. ( A ) Cells were infected with Wuhan variant (500 pfu/well) or ( B ) Delta variant (500 pfu/well) for 2 h and 24 h in absence (untreated) and presence of TAT-I24 (10 µM) or TAT (10 µM), followed by staining for dsRNA (FITC/green); nuclei were stained with DAPI (blue), and one representative section is shown. Scale bars indicate 50 µm. ( C ) Quantification of dsRNA positive signals (FITC/green) from three randomly chosen sections normalized against DAPI (blue) signals and expressed as % of control; mean + SD from dsRNA relative to untreated controls are shown. Significant differences were detected using one-way ANOVA (Kruskal–Wallis multiple comparison of means); p -value: p ≤ 0.05 (*).

Article Snippet: After washing three times, a secondary fluorescent-labeled antibody (Alexa Fluor 488 goat-anti mouse; #A-11008, Invitrogen/Thermofisher; Waltham, MA, USA) was diluted 1:400 in 1% BSA and the coverslips were incubated in the dark for 1 h. After washing, the coverslips were carefully lifted from the cell culture plates, flipped in reverse, and placed on a glass slide spotted with 20 μL of ProLong ® Gold Antifade Mountant with DNA stain 4′,6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI) (#P36931, Invitrogen/Thermofisher, Waltham, MA, USA;).

Techniques: Infection, Variant Assay, Staining, Control, Comparison

Journal: Viruses

Article Title: The Virus Entry Pathway Determines Sensitivity to the Antiviral Peptide TAT-I24

doi: 10.3390/v17040458

Figure Lengend Snippet: Localization of TAT-I24 in endosomal compartments. ( A ) Localization of Rho-TAT-I24 (red) in COS-7 cells after 15, 90, and 180 min incubation or ( B ) in Vero E6 cells after 30 and 120 min incubation. ( C ) Localization of Rho-TAT-I24 after 120 min either alone or in presence of 30 µM of CQ. ( D ) Staining of COS-7 cells for LAMP1 (green) after 30 min of incubation with Rho-TAT-I24 (red). Arrows indicate co-localization of Rho-TAT-I24 and LAMP1. ( E ) Overexpression of GFP-Rab5 (green) and lack of co-localization with Rho-TAT-I24 (red), or ( F ) overexpression of GFP-Rab14 (green) and incubation with Rho-TAT-I24 (red). Arrows indicate co-localization of GFP-Rab14 and Rho-TAT-I24 in enlarged vesicles. Nuclei are stained with DAPI (blue). Scale bars indicate 40 µm. ( G ) Co-localization of GFP-Rab5 or GFP-Rab14 with Rho-TAT-I24, expressed as percent (%) of total GFP signal, was determined from four areas of two independent experiments using color threshold tool in ImageJ.

Article Snippet: After washing three times, a secondary fluorescent-labeled antibody (Alexa Fluor 488 goat-anti mouse; #A-11008, Invitrogen/Thermofisher; Waltham, MA, USA) was diluted 1:400 in 1% BSA and the coverslips were incubated in the dark for 1 h. After washing, the coverslips were carefully lifted from the cell culture plates, flipped in reverse, and placed on a glass slide spotted with 20 μL of ProLong ® Gold Antifade Mountant with DNA stain 4′,6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI) (#P36931, Invitrogen/Thermofisher, Waltham, MA, USA;).

Techniques: Incubation, Staining, Over Expression