anti fade mounting medium with dapi  (Vector Laboratories)

Journal: Materials Today Bio

Article Title: 4D fabrication of scaffolds facilitates the construction of cholangiocyte monolayers from human and mouse liver derived organoids

doi: 10.1016/j.mtbio.2025.102757

Figure Lengend Snippet: PT-PLATMC@PDA scaffold supports the survival, proliferation and spreading of cholangiocytes to form a monolayer. Cell viability (A) and density (B) of cholangiocytes seeded on PT-PLATMC@PDA films in culture for 1 day, 3 days, and 7 days (n = 3). Fluorescence microscope images of mouse liver derived cholangiocyte organoid seeded on the PT-PLATMC@PDA film in culture for 7 days (C), 14 days (D), 21 days (E) and 35 days (F) stained with DAPI, ZO-1, and CK19. DAPI and ECAD staining of the 21-day cultured mouse cholangiocytes (G) and 37-day cultured human cholangiocytes (H) seeded SMP PT-PLATMC@PDA planar films.

Article Snippet: The samples were washed with PBS, mounted with 100 μl of anti-fade mounting medium with DAPI (Vector Laboratories Inc, USA), and analyzed under a confocal laser scanning microscope (Zeiss LSM880 with Ariyscan, Germany).

Techniques: Fluorescence, Microscopy, Derivative Assay, Staining, Cell Culture

Journal: Materials Today Bio

Article Title: 4D fabrication of scaffolds facilitates the construction of cholangiocyte monolayers from human and mouse liver derived organoids

doi: 10.1016/j.mtbio.2025.102757

Figure Lengend Snippet: Construction of a single-layer bile duct epithelium on PT-PLATMC@PDA tubular scaffolds. PT-PLATMC@PDA tubes seeded with (A) mouse derived cholangiocyte organoids cultured for 21 days and (B) human derived cholangiocyte organoids cultured for 37 days immuno-stained for DAPI, ZO-1and/or F Actin.

Article Snippet: The samples were washed with PBS, mounted with 100 μl of anti-fade mounting medium with DAPI (Vector Laboratories Inc, USA), and analyzed under a confocal laser scanning microscope (Zeiss LSM880 with Ariyscan, Germany).

Techniques: Derivative Assay, Cell Culture, Staining

Journal: bioRxiv

Article Title: Satellite Glial Cells Control Sensory Neuron Excitability via the Release of Fibulin-2

doi: 10.64898/2026.02.13.705760

Figure Lengend Snippet: A. Representative images of DRG sections from WT and Fibulin-2 KO mice immunostained for Fibulin-2, FABP7, TUJ1, and DAPI. Scale bar 50μm. Two DRG sections from n=3 mice were imaged for each group. B. Super-resolution imaging of DRG tissue section highlighting the subcellular localization of Fibulin-2 in SGCs near the perinuclear region. Fibulin-2 (Red), FABP7 (White), TUJ1 (Green), and DAPI (Blue). Scale bar 2μm C. Electron micrograph of DRG sections showing the portion of an SGC covering the neuron soma. The SGC cytoplasm contains multivesicular bodies (white arrow), vesicle fusion/budding at the plasma membrane (red arrow), and a Golgi apparatus (asterisk). Scale bar 1μm D. Immunofluorescence of DRG section showing localization of Fibulin-2 in SGCs (labeled for Fabp7 or PDPN) surrounding neurons labeled with NeuN, NF200, TRPV1, TH, TRKA, and IB4. Scale bar 50μm. E. Quantification of the size distribution of the indicated DRG neurons subtypes. The size of neurons surrounded by Fibulin-2 positive SGC is indicated by a red dotted line. A total of 8956 neurons were analyzed from n=7 mice, with three DRG sections per mice. F. Quantification of the percentage of neuronal subtype surrounded by Fibulin-2-positive SGCs. Each data point in the bar graph represents an independent biological replicate. NF200 (n=5 mice), TRPV1 (n=5 mice), TRKA (n=3 mice), IB4 (n=4 mice), TH (n=4). Two sections from each biological replicate were imaged and used for quantification. Error bars represent SEM.

Article Snippet: Sections were mounted and coverslipped using VECTASHIELD anti-fade mounting media with DAPI (Vector Laboratory, Cat# H-2000).

Techniques: Imaging, Clinical Proteomics, Membrane, Immunofluorescence, Labeling

Journal: bioRxiv

Article Title: Satellite Glial Cells Control Sensory Neuron Excitability via the Release of Fibulin-2

doi: 10.64898/2026.02.13.705760

Figure Lengend Snippet: A . Voltage protocols for measurement of different types of K + currents: total ( I Total ), K-type ( I K ) and A-type ( I A ) K + currents. B . Sample traces of voltage-dependent K + currents I total (left), I K (middle) and I A (right) evoked by the protocols in ( A ) from Control (upper panel) and rFibulin-2 treated DRG cells (lower panel). C . rFibulin-2 increases voltage-dependent K + currents I Total (left), I K (middle) and I A (right) in DRG cells. Insert bar graphs are K + currents at membrane potential of -10 mV (around voltage threshold level), indicating that rFibulin-2 decreases excitability mainly mediated by enhancement of I A conductance, which reduces input resistance. Number of cells tested from 3 independent experiments: control n = 10; rFibulin-2: n = 8. D . Phrixotoxin-1 (PaTx1) was used to isolate Kv4 current evoked by voltage ramp (-100 to +20 mV, 100 mV/s). Sample traces of ramp-evoked K + currents before (a) and during (b) application of PaTx1, and the PaTx1-sensitive current (c, c = a - b). Currents were normalized to membrane capacitance for better comparison. E . I-V curves were constructed from the ramp-evoked Kv4 current (mean current value over 0.1 mV intervals from averages of five trials for each cell to approximate quasi-steady-state current). Note PaTx1 significantly increases the Kv4 current when the membrane potentials are depolarized to positive values greater than -25 mV. Number of cells tested from 3 independent experiments: control n = 6; rFibulin-2: n = 6; T-test; * P < 0.05; ** P < 0.01. F . Representative western blot of control and Fibulin-2 KO DRG lysate analyzed for Fibulin-2 and Kv4.2. GAPDH is used as a loading control. G . Quantification of Kv4.2 expression in control and Fibulin-2 KO mice. n=3 WT and n=3 Fibulin-2 KO mice. T-test; ** P < 0.01. H . Representative western blot of control and Fibulin-2 KO DRG lysate analyzed for Fibulin-2 and Kv4.3. GAPDH is used as a loading control. I . Quantification of Kv4.3 expression in control and Fibulin-2 KO mice. n=3 WT and n=3 Fibulin-2 KO mice. T-test; *** P < 0.001 J . Fibulin-2 KO mice show hypersensitivity to mechanical stimuli compared to controls, measured by the Von Frey Test. 12 WT and 8 Fibulin-2 KO mice were used. Two-Way Anova. ∗p < 0.05, ∗∗p < 0.01, ***p<0.001. K . Fibulin-2 KO mice exhibit hypersensitivity to heat stimuli compared to controls, measured by the Hot-Plate test. 12 WT and 8 Fibulin-2 KO mice were used. Two-Way Anova. ∗p < 0.05, ∗∗p < 0.01, ***p<0.001. L . Fibulin-2 KO mice exhibit hypersensitivity to cold stimuli compared to controls, measured by the Cold-Plate test. 12 WT and 8 Fibulin-2 KO mice were used. Two-Way Anova. ∗p < 0.05, ∗∗p < 0.01, ***p<0.001. M . Representative immunofluorescence images of the hindpaw of control and Fibulin-2 KO mice immunostained for PGP9.5 (white) and DAPI (blue). Three sections from n=3 mouse per group were used. N . Quantification of intraepidermal nerve fiber density (IENFD). n=3 mice per genotype. T-test, ns- non-significant

Article Snippet: Sections were mounted and coverslipped using VECTASHIELD anti-fade mounting media with DAPI (Vector Laboratory, Cat# H-2000).

Techniques: Control, Membrane, Comparison, Construct, Western Blot, Expressing, Hot Plate Test, Immunofluorescence

Journal: Scientific Reports

Article Title: Targeting cell surface GRP78-CD44v interaction suppresses cell migration in triple-negative breast cancer cells

doi: 10.1038/s41598-025-33441-5

Figure Lengend Snippet: Cell surface GRP78 co-localizes with CD44v in vitro and in vivo in MDA-MB-231 breast cancer cells. ( a ) Left: Immunofluorescence and compressed z-stack confocal image of a non-permeabilized MDA-MB-231 cell showing cell surface GRP78 (red) and CD44v (green); nucleus stained with DAPI (blue). The arrow indicates the direction of cell migration. Ante, anterior; Mid, middle; Post, posterior. Scale bar, 20 μm. Middle: Co-localization of GRP78 and CD44v (magenta) at the anterior region overlaid on DIC image using FIJI-ImageJ. Right: Quantification of csGRP78, csCD44v, and co-localized signals across different regions of unipolar cells. Each dot represents one cell. Statistics: unpaired two-tailed Student’s t-test; error bars: standard error of the mean (SEM). ( b – d ) Flow cytometry analysis of csGRP78 and csCD44v expression on live MDA-MB-231 cells. Unpaired two-tailed Student’s t-test was used to calculate the p -values. ( b ) CD44v + cells (magenta, n = 23,820); CD44v -/low cells (green, n = 1,629). ( c ) Box-and-whisker plot of csGRP78 expression in CD44v + vs. CD44v -/low cells. “x” indicates the mean; dots, outliers. ( d ) Box-and-whisker plot of side scatter area (SSC-A) and forward scatter area (FSC-A) in subpopulations: CD44v + GRP78 + (dark magenta, n = 3,600), CD44v + GRP78 - (light magenta, n = 4,087), CD44v -/low GRP78 + (dark green, n = 25), CD44v -/low GRP78 - (light green, n = 1,663). “x” indicates the mean; dots, outliers. ( e ) Confocal images of GRP78 (red) and CD44v (green) in MDA-MB-231 tumor xenografts. Nuclei stained with DAPI (blue). The boxed region indicates the enlarged area shown in the lower panels. Arrows indicate co-localization. The thickness of the image section: 0.3 μm. Scale bars, 20 μm; 5 μm (enlarged images). ( f ) Western blot analysis of whole cell lysates from non-treated (NT) MDA-MB-231 cells and cells treated with 76-E6 antibody or control IgG for 24 h. Numbers below bands represent relative protein levels normalized to GAPDH or total Src. Magenta arrows indicate lower-molecular-weight protein forms. CD44v and GAPDH were run in the same gel while pSrc and tSrc were run in parallel on a different gel. The whole cell lysates used in the Western blot analysis were prepared from the same experiment with two biological replicates. The nitrocellulose membrane was separated into different pieces according to the molecular marker in order to blot multiple proteins. The full-length images are available in Figs. S4 and S5 of Supplementary Information 2. The raw statistical data are provided in Supplementary Information 1.

Article Snippet: The next day, cells were incubated with an AlexaFluor-488 secondary antibody (Thermo Scientific, Waltham, MA) at RT for 1 h. Coverslips were then mounted using Vectashield anti-fade medium with DAPI (Vector Laboratories, Burlingame, CA).

Techniques: In Vitro, In Vivo, Immunofluorescence, Staining, Migration, Two Tailed Test, Flow Cytometry, Expressing, Whisker Assay, Western Blot, Control, Molecular Weight, Membrane, Marker