immunohistochemical staining  (Proteintech)

Journal: Biochemistry and Biophysics Reports

Article Title: Oxysterol-binding protein-related protein 6 regulates neuronal morphology and migration of cerebellar granule cells during cerebellar development in vivo

doi: 10.1016/j.bbrep.2026.102585

Figure Lengend Snippet: ORP6 RNAi decreases cell motility of primary cultured cerebellar granule cells (CGCs). Cell-tracking images of primary cultured CGCs transfected with control RNA (A) or ORP6 RNAi (B), stained with Hoechst. (C) The accumulated distance of primary cultured CGCs transfected with control or ORP6 RNAi is automatically analyzed by PerkinElmer Harmony 4.9 Image Analysis Software. All colored arrows indicate the distance and direction of cell movement. Data are collected from five independent cell culture preparations, and the accumulated distance of each two groups is shown as the mean ± SE. Statistical analysis is performed using Welch's t -test. A P value less than 0.05 is considered statistically significant. Bars, 50 μm.

Article Snippet: Neuro-2A cells, primary cultured CGCs, and cerebellar sections were incubated with primary antibodies at 4 °C overnight, followed by incubation with secondary antibodies at 37 °C for 1 h, as described in , Cells and cerebellar sections were then washed four times with PBS and incubated with Hoechst stain (346-07951, DOJINDO, Kumamoto, Japan) in PBS at RT for 10 min. After washing with PBS, the cerebellar sections were mounted with CC/Mount (K002, Diagnostic Biosystems, Pleasanton, CA, USA).

Techniques: Cell Culture, Cell Tracking Assay, Transfection, Control, Staining, Software

Journal: Biochemistry and Biophysics Reports

Article Title: Oxysterol-binding protein-related protein 6 regulates neuronal morphology and migration of cerebellar granule cells during cerebellar development in vivo

doi: 10.1016/j.bbrep.2026.102585

Figure Lengend Snippet: ORP6 int impaired the migration of cerebellar granule cells (CGCs) in the developing cerebellum. (A) Experimental design of gene transfection into P7 mice cerebellum by in vivo electroporation and tissue collection. Sagittal section of P9 cerebellum transfected with pCAGGS-AcGFP-C (B–D) or pCAGGS-AcGFP-C-ORP6 int (E–G), and immunostained with anti-calbindin antibody (C and F). The cerebellar laminar structure is identified as follows: the calbindin-positive Purkinje cell layer (PCL) and molecular layer (ML), which lies superficial to the PCL and contains sparsely Hoechst-stained nuclei. The external granular layer is the outermost layer of the ML, a region with dense Hoechst-stained nuclei, and the internal granular layer located beneath the calbindin-positive PCL. Arrows indicate distribution of CGCs expressing pCAGGS-AcGFP-C or pCAGGS-AcGFP-C-ORP6 int. Ratio of cells transfected with pCAGGS-AcGFP-C (H) or pCAGGS-AcGFP-C-ORP6 int (I) in each layer to total cells. Data are collected from four animals, and the cell number of each two groups is shown as the mean ± SE. Statistical analysis is performed using Welch's t -test. A P value less than 0.05 is considered statistically significant. Bars, 50 μm.

Article Snippet: Neuro-2A cells, primary cultured CGCs, and cerebellar sections were incubated with primary antibodies at 4 °C overnight, followed by incubation with secondary antibodies at 37 °C for 1 h, as described in , Cells and cerebellar sections were then washed four times with PBS and incubated with Hoechst stain (346-07951, DOJINDO, Kumamoto, Japan) in PBS at RT for 10 min. After washing with PBS, the cerebellar sections were mounted with CC/Mount (K002, Diagnostic Biosystems, Pleasanton, CA, USA).

Techniques: Migration, Transfection, In Vivo, Electroporation, Staining, Expressing

Journal: Bioactive Materials

Article Title: Skin-mimetic bilayer hydrogel normalizes diabetic wound healing by orchestrating inflammatory cell dynamics: An early intervention strategy

doi: 10.1016/j.bioactmat.2026.02.025

Figure Lengend Snippet: Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining for Ly-6G in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.

Article Snippet: The infiltration of pro-inflammatory (M1) macrophages and polarization of M2c macrophages were analyzed by immunofluorescence staining using antibodies against iNOS (Servicebio, GB11119) and CD163 (Servicebio, GB14027), respectively.

Techniques: Immunohistochemical staining, Staining, Saline, Expressing, Immunofluorescence, Generated, Standard Deviation

Journal: Bioactive Materials

Article Title: Skin-mimetic bilayer hydrogel normalizes diabetic wound healing by orchestrating inflammatory cell dynamics: An early intervention strategy

doi: 10.1016/j.bioactmat.2026.02.025

Figure Lengend Snippet: Bilayer hydrogel modulates M2c macrophage polarization in the later healing phase of diabetic wound. a Experimental timeline for assay of M2c macrophage polarization. b Immunofluorescence staining for CD163 in wounds on days 3, 6, 9 and 12 after injury. c Quantitative analysis of CD163 + cells in each group. d-f Relative expression of MerTK, IL-10, and TGF-β1 on day 6. g Schematic illustration of M2c macrophage polarization regulated by SP/IL-10@Bilayer and its contribution to inflammation resolution. All data were generated from three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05 and ##p < 0.01; ∗ means significant difference compared to model group. ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001; & means significant difference compared to SP/IL-10@Bilayer group. & p < 0.05.

Article Snippet: The infiltration of pro-inflammatory (M1) macrophages and polarization of M2c macrophages were analyzed by immunofluorescence staining using antibodies against iNOS (Servicebio, GB11119) and CD163 (Servicebio, GB14027), respectively.

Techniques: Immunofluorescence, Staining, Expressing, Generated, Standard Deviation

Journal: International Journal of Molecular Medicine

Article Title: Novel Hsp90 inhibitor JD-02 inhibits HSV-1 infection via the Raf/MEK/ERK signaling pathway

doi: 10.3892/ijmm.2026.5810

Figure Lengend Snippet: JD-02 inhibits HSV-1 early infection. (A) Schematic representation of the time-of-addition assay. (B) Western blot analysis of viral proteins (gB, ICP0, gD and ICP27) from HaCaT cells infected with HSV-1 and treated with JD-02 (0.5 μ M) for indicated time. (C) The DNA copy number of UL54 from HaCaT cells with HSV-1 infection and treated with JD-02. (D-F) Viral plaque assay detecting JD-02 effect on (D) viral inactivation, (E) viral attachment and (F) viral penetration. (G) Immunofluorescence analysis of the distribution of the viral protein VP5 (green) within the cytoplasm and nucleus (DAPI, blue). Scale bar, 10 μ m. Data are presented as the mean ± SD (n=3). * P<0.05 and ** P<0.01 compared with the HSV-1 group. HSV, Herpes Simplex Virus; ns, not significant.

Article Snippet: The cells to be treated were fixed with 4% paraformaldehyde (PFA), penetrated with 0.1% NP-40 for 4 min, and then blocked with 5% bovine serum albumin (BSA) for 90 min before staining with VP5 (1:100; cat. no. sc-13525; Santa Cruz Biotechnology, Inc.) overnight at 4°C.

Techniques: Infection, Western Blot, Viral Plaque Assay, Immunofluorescence, Virus