vectashield plus antifade 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: Neural Regeneration Research

Article Title: Injury-induced KIF4A neural expression and its role in Schwann cell proliferation suggest a dual function for this kinesin in neural regeneration

doi: 10.4103/NRR.NRR-D-24-00232

Figure Lengend Snippet: Immunohistochemistry analysis of KIF4A expression in the uninjured spinal cord. Fluorescence microphotographs of 50 µm sagittal sections of uninjured rat spinal cord, where KIF4A (red) is detected in cell bodies of GFAP-negative neurons of the gray matter (arrowheads) and axon-like structures of the white matter (arrows) (A); in axons, where it co-labels with neurofilament (NF) 160/200 (arrows) (B); and in myelinated axons of the white matter (juxtaposed labeling with MBP) (C). (D) KIF4A (red) staining is absent from GFAP-positive glial structures (arrowheads). (E) Vesicular acetylcholine transporter (VAChT)-positive motor neurons of the ventral white matter (arrows). KIF4A also stains smaller VAChT-negative cells (asterisks) but is apparently absent from VAChT-positive axonal terminals synapsing with some of these spinal motor neurons (arrowheads). The dotted box surrounds an area presented with a higher magnification on the right. Images from the same panel (different channels of the same image view) share the same scale bar. DAPI: 4′,6-Diamidino-2-phenylindole; GFAP: glial fibrillary acidic protein; MBP: myelin basic protein.

Article Snippet: Nuclei were counterstained with VECTASHIELD® Antifade Mounting Medium with DAPI (Vector Laboratories, H-1200) or Hoechst 33258 (09460, PolySciences, Warrington, PA, USA).

Techniques: Immunohistochemistry, Expressing, Fluorescence, Labeling, Staining

Journal: Neural Regeneration Research

Article Title: Injury-induced KIF4A neural expression and its role in Schwann cell proliferation suggest a dual function for this kinesin in neural regeneration

doi: 10.4103/NRR.NRR-D-24-00232

Figure Lengend Snippet: KIF4A expression in cross sections of uninjured rat DRG. Fluorescence immunohistochemistry analysis using IB4 dye (green) to stain the smaller neurons in DRG (A) and anti-NF200 antibody (green) (B) to stain the larger DRG neurons. KIF4A (red) mainly labels some nuclei of both types of neurons (arrows in high magnification images, with faint blue staining), and also satellite cells surrounding the neurons (arrowheads). Some faint cytoplasmic labeling is sometimes observed (e.g., high magnification inset, A). The brightness in DAPI channel was intensified to reveal the less intense neuronal nuclei, and images from the same panel (different channels of the same image view) share the same scale bar. The dotted boxes surround areas presented with a higher magnification. The dotted circles identify the same DRG neurons in the different channels of a same image. DAPI: 4′,6-Diamidino-2-phenylindole; DRG: dorsal root ganglia; NF: neurofilament.

Article Snippet: Nuclei were counterstained with VECTASHIELD® Antifade Mounting Medium with DAPI (Vector Laboratories, H-1200) or Hoechst 33258 (09460, PolySciences, Warrington, PA, USA).

Techniques: Expressing, Fluorescence, Immunohistochemistry, Staining, Labeling

Journal: Neural Regeneration Research

Article Title: Injury-induced KIF4A neural expression and its role in Schwann cell proliferation suggest a dual function for this kinesin in neural regeneration

doi: 10.4103/NRR.NRR-D-24-00232

Figure Lengend Snippet: KIF4A protein expression in rat uninjured peripheral nerves. Fluorescence immunohistochemistry analyses of rat sciatic nerve cross-sections (A and C) and longitudinal sections (B and D). (A and B) KIF4A (immunostained in red) co-labels with NF 160/200 (green), being highly abundant in the nerve axons (arrows). Extra-axonal KIF4A staining is also visible in A, surrounding the axons (arrowheads), suggesting that in peripheral nerves KIF4A is also expressed by Schwann cells. (C and D) Confirming that hypothesis, extra-axonal KIF4A co-labels with the Schwann cells’ S100 marker (arrows), and is observed in both cytoplasm and nuclei. KIF4A also stains what appear to be unmyelinated axonal fibers (arrowheads in C). The dotted boxes surround areas presented with a higher magnification. Images from the same panel (different channels of the same image view) share the same scale bar. DAPI: 4′,6-Diamidino-2-phenylindole; NF: neurofilament.

Article Snippet: Nuclei were counterstained with VECTASHIELD® Antifade Mounting Medium with DAPI (Vector Laboratories, H-1200) or Hoechst 33258 (09460, PolySciences, Warrington, PA, USA).

Techniques: Expressing, Fluorescence, Immunohistochemistry, Staining, Marker

Journal: Neural Regeneration Research

Article Title: Injury-induced KIF4A neural expression and its role in Schwann cell proliferation suggest a dual function for this kinesin in neural regeneration

doi: 10.4103/NRR.NRR-D-24-00232

Figure Lengend Snippet: KIF4A protein expression in human peripheral nerves. KIF4A chromogenic (A, B) and fluorescence (C–F) immunostainings of cross-sections of a human craniofacial nerve. (A–D) The same axonal pattern, previously observed in rat nerve, is evident (exemplified by some asterisks in A–F); some longitudinal portions of myelinated axons (arrows in B and D) are also visible within the transversal sections, along with axonal bundles of apparent unmyelinated fibers (arrowheads in C and F). (E and F) Most of the KIF4A extra-axonal staining in nerves occurs in Schwann cells, since KIF4A staining is mainly visible either in axons (asterisks) or in the cytoplasm of S100-positive cells ensheathing these axons (arrows). The dotted boxes surround areas presented with a higher magnification. Images from the same panel (different channels of the same image view) share the same scale bar. DAPI: 4′,6-Diamidino-2-phenylindole.

Article Snippet: Nuclei were counterstained with VECTASHIELD® Antifade Mounting Medium with DAPI (Vector Laboratories, H-1200) or Hoechst 33258 (09460, PolySciences, Warrington, PA, USA).

Techniques: Expressing, Fluorescence, Staining

Journal: Neural Regeneration Research

Article Title: Injury-induced KIF4A neural expression and its role in Schwann cell proliferation suggest a dual function for this kinesin in neural regeneration

doi: 10.4103/NRR.NRR-D-24-00232

Figure Lengend Snippet: KIF4A expression in DRG neurons following peripheral nerve injury. (A) Real-time qPCR analysis of Kif4a mRNA levels in rat sensory DRG neurons at various time points following sciatic nerve crush or transection (with suture). Values are presented as mean ± SD fold increases over mRNA levels of control time-matched sham-operated animals, collected at the same time points as each injury sample and taken as ‘1’ for each time point; n = 3. (*) crush vs. time-matched sham data; (#) transection-with-suture vs. time-matched sham data; # P < 0.05; ** P < 0.01, using unpaired t -test with Welch correction. (B) Fluorescence KIF4A and IB4 staining in a cross-section of DRG from a sham-operated rat, showing KIF4A at the neurons’ cytoplasm (asterisk), plasma membrane, and in some nuclei (arrows). (C, D) Fluorescence KIF4A immunostaining in DRG cross-sections at 2 dpi following nerve crush (C) or transection with suture (D). Neurons were labeled with IB4 (C) or NF200 (D) (green). An increased KIF4A staining (red) is apparent at the neurons’ cytoplasm, membrane, and nuclei. Nuclear staining is more visible in the crush condition (C, arrows for IB4 + and arrowheads for IB4 – neurons) than in the transected condition (D, arrows). Of note, the intensity of the DAPI channel was increased to allow visualization of the less intense neuronal nuclei. The dotted boxes surround areas presented with a higher magnification. Images from the same panel (different channels of the same image view) share the same scale bar. DAPI: 4′,6-Diamidino-2-phenylindole; DRG: dorsal root ganglia; NF: neurofilament.

Article Snippet: Nuclei were counterstained with VECTASHIELD® Antifade Mounting Medium with DAPI (Vector Laboratories, H-1200) or Hoechst 33258 (09460, PolySciences, Warrington, PA, USA).

Techniques: Expressing, Control, Fluorescence, Staining, Clinical Proteomics, Membrane, Immunostaining, Labeling

Journal: Neural Regeneration Research

Article Title: Injury-induced KIF4A neural expression and its role in Schwann cell proliferation suggest a dual function for this kinesin in neural regeneration

doi: 10.4103/NRR.NRR-D-24-00232

Figure Lengend Snippet: KIF4A mRNA and protein expression in the distal stump of an injured nerve. (A) Real-time quantitative PCR analysis of Kif4a mRNA levels in the distal stump of the sciatic nerve at various time points following nerve lesion by crush or transection with suture. Values are presented as mean ± SD fold increases over mRNA levels of control time-matched sham-operated animals, collected at the same time points as each injury sample and taken as ‘1’ for each time point; n = 5. (*) crush vs. time-matched sham data; (#) transection-with-suture vs. time-matched sham data; */# P < 0.05, ## P < 0.01, by unpaired t -test with Welch correction. (B–D) Fluorescence immunohistochemistry analyses of longitudinal sections of sciatic nerve distal stumps, either uninjured (B) or injured (C, D) and collected at 7 dpi. (B) In sham-operated nerves, KIF4A (in red) is observed in S100-positive Schwann cells (in green, arrow), and in S100-negative axons that are Schwann cells-ensheathed (arrowhead). (C) In the transected and sutured nerve collected at 7 dpi, a high number of KIF4A/S100-positive Schwann cells is observed. (D) In a crushed nerve collected at 7 dpi, a high number of cells double-stained for KIF4A (red) and PDGFRα (green, glial cells marker) is also observed, supporting that KIF4A is up-regulated in Schwann cells (arrows) following injury. KIF4A is also observed in other, PDGFRα-negative, cells (asterisk). The dotted boxes surround areas presented with a higher magnification. Images from the same panel (different channels of the same image view) share the same scale bar. DAPI: 4′,6-Diamidino-2-phenylindole; dpi: days post-injury; PDGFRα: platelet-derived growth factor receptor alpha.

Article Snippet: Nuclei were counterstained with VECTASHIELD® Antifade Mounting Medium with DAPI (Vector Laboratories, H-1200) or Hoechst 33258 (09460, PolySciences, Warrington, PA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Fluorescence, Immunohistochemistry, Staining, Marker, Derivative Assay

Journal: Neural Regeneration Research

Article Title: Injury-induced KIF4A neural expression and its role in Schwann cell proliferation suggest a dual function for this kinesin in neural regeneration

doi: 10.4103/NRR.NRR-D-24-00232

Figure Lengend Snippet: KIF4A expression in rat regenerating sciatic nerve fascicles, upon nerve transection with conduit implantation. (A) Schematic image of the conduit “Neuragen® Nerve Guide” (adapted from Integra LifeSciences Corporation). (B–D) Microphotographs evidencing KIF4A distribution (red) in a transected sciatic nerve repaired with the NeuraGen conduit, collected at 10 dpi. (B) Colorimetric immunohistochemical detection of the axonal growth-cone marker GAP43 (left) and KIF4A (right) in adjacent tissue sections, inside the conduit; a positive reaction is observed in all the regenerating nerve tissue, but particularly more intense in the left half of the regenerating nerve (high magnification images). (C) Immunofluorescence analysis of KIF4A (red) and GAP43 (green) in the conduit-supported regenerating nerve, confirming KIF4A staining in GAP43-marked regrowing axons (arrows). (D) In this condition, KIF4A (red) is also expressed in the repair supporting Schwann cells (green S100-positive cells; arrowheads in C and high magnification images in D). The dotted boxes surround areas presented with a higher magnification. Images from the same panel (different channels of the same image view) share the same scale bar. DAPI: 4′,6-Diamidino-2-phenylindole; GAP43: growth-associated protein 43.

Article Snippet: Nuclei were counterstained with VECTASHIELD® Antifade Mounting Medium with DAPI (Vector Laboratories, H-1200) or Hoechst 33258 (09460, PolySciences, Warrington, PA, USA).

Techniques: Expressing, Immunohistochemical staining, Marker, Immunofluorescence, Staining

Journal: Neural Regeneration Research

Article Title: Injury-induced KIF4A neural expression and its role in Schwann cell proliferation suggest a dual function for this kinesin in neural regeneration

doi: 10.4103/NRR.NRR-D-24-00232

Figure Lengend Snippet: KIF4A role in the proliferation of immature Schwann cells. (A) Kif4a expression was analyzed by real-time quantitative PCR in rat neonatal Schwann cells cultured in non-proliferative (confluent) and proliferative (sub-confluent) conditions. Values are presented as fold increases over mRNA levels in the non-proliferative condition, taken as “1”; mean ± SD, n = 3. (B, C) Schwann cells cultured under proliferative conditions were transfected for 48 hours with an anti-Kif4a shRNA GFP-expressing plasmid (“shKif4”) or the empty plasmid (only GFP-expressing, “Ctrl”). Arrows indicate green cells, transfected with the empty plasmid (“Ctrl” panel) or with the anti-Kif4a shRNA (‘shKif4’ panel); arrowheads indicate neighbor non-transfected cells. (B) Left: When transfected with the shKif4 plasmid, the KIF4A immunostaining (red) highly decreases (arrow in “shKif4” panel). Right: Downregulation of Kif4a mRNA in the Schwann cells population was confirmed by real-time qPCR; mean fold ± SD values, presented as fold increases over mRNA levels in the control levels, taken as ‘1’ ( n = 3). (C) Left: The percentage of proliferating cells, exhibiting red Ki67/blue DAPI-positive nuclei, was scored in the green fluorescing cells population (arrows). Right: results are graphically expressed as a percentage of Ki67-positive/GFP-positive cells in the GFP green fluorescing population (mean ± SD; n = 5–6, where 200–500 GFP-expressing cells were scored per ‘ n ’). * P < 0.05, *** P < 0.001, using unpaired t -test with Welch correction. Images from the same panel (different channels of the same image view) share the same scale bar. Ctrl: Conytol; DAPI: 4′,6-diamidino-2-phenylindole; GFP: green fluorescent protein; SCs: Schwann cells.

Article Snippet: Nuclei were counterstained with VECTASHIELD® Antifade Mounting Medium with DAPI (Vector Laboratories, H-1200) or Hoechst 33258 (09460, PolySciences, Warrington, PA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Cell Culture, Transfection, shRNA, Plasmid Preparation, Immunostaining, Control

Journal: Atherosclerosis Plus

Article Title: Heterozygous transferrin receptor 1 deletion reduces atherosclerotic lesion formation in apolipoprotein E-deficient mice

doi: 10.1016/j.athplu.2025.12.003

Figure Lengend Snippet: Effects of heterozygous transferrin receptor 1 deletion on atherosclerotic lesion formation in apolipoprotein E-deficient mice . (A) Experimental protocol; 6 weeks-old ApoE −/− or ApoE −/− /TfR1 +/− mice were fed for 16 weeks with a high-fat diet (HFD), and representative photographs of Oil Red O-stained sections (scale bar: 500 μm) from the aortic sinus and en face whole aorta, and quantification of Oil Red O-stained area in the aortic sinus sections and en face whole aortas of ApoE −/− and ApoE −/− /TfR1 +/− mice at 16 weeks after a high-fat diet (HFD) ( n = 6–10/group). (B) Representative images of TfR1, 4-hydroxynonenal (4HNE), and CD68 staining in the aortic sinus sections (scale bar: 500 μm) in ApoE −/− and ApoE −/− /TfR1 +/− mice at 16 weeks after a high-fat diet (HFD). (C) Quantification of TfR1, 4HNE, and CD68-positive area in the aortic sinus sections of ApoE −/− and ApoE −/− /TfR1 +/− mice at 16 weeks after a high-fat diet (HFD) ( n = 6/group). (D) Representative merged image of immunofluorescence staining of CD68, TfR1, and DAPI in the aortic sinus sections (scale bar: upper-left 500, upper-right and lower-left 200, lower-right 100 μm) of ApoE −/− mice at 16 weeks after a high-fat diet. DAPI, 4′,6-diamidino-2-phenylindol. (E) Western blot analysis for aortic TfR1, transferrin, and ferritin in ApoE −/− and ApoE −/− /TfR1 +/− mice at 16 weeks after a high-fat diet (HFD) ( n = 4/group). (F) Representative images of Oil Red O, TfR1, 4HNE, and CD68 staining in the aorta sections (scale bar: 100 μm) in ApoE −/− and ApoE −/− /TfR1 +/− mice at 16 weeks after a high-fat diet (HFD). (G) Quantification of Oil Red O, TfR1, 4HNE, and CD68-positive area in the aorta sections of ApoE −/− and ApoE −/− /TfR1 +/− mice at 16 weeks after a high-fat diet (HFD) ( n = 6/group). (H) Representative images of immunofluorescence staining of CD68 and TfR1, and merged image of CD68, TfR1, and DAPI in the aortic sections (scale bar: upper-left, upper-right, and lower-left 100, lower-right 50 μm) of ApoE −/− mice at 16 weeks after a high-fat diet. (I) Body weight, serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL), high-density lipoprotein cholesterol (HDL), triglycerides (TG), iron, hemoglobin, and mean corpuscular volume (MCV) in ApoE −/− and ApoE −/− /TfR1 +/− mice at 16 weeks after a high-fat diet (HFD). N = 5–6/group. Results are shown as means ± SEM. Student's t-test was used and a probability value < 0.05 was described in each Figure. ns, not significant.

Article Snippet: Nuclei were counterstained with 4’,6-diamidino-2-phenylindol (catalog # H-1800 , Vector Laboratories).

Techniques: Staining, Immunofluorescence, Western Blot