hoechst stain  (Cytoskeleton Inc)

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: Journal of Advanced Research

Article Title: HDAC2 enhances the antimicrobial activity of neutrophils by promoting the formation of neutrophil extracellular traps (NETs) in sepsis

doi: 10.1016/j.jare.2025.08.041

Figure Lengend Snippet: HDAC2 promotes NETs formation through the acetylation-citrullination pathway. (A–C) Representative images for LPS (50 μg/ml)-stimulated primary neutrophils from HDAC2 WT and HDAC2 KO mice. (A) Immunostaining for histone H3K18 acetylation (green) was performed with DNA counterstained with Hoechst 33342 (blue) and corresponding MFI of quantitative data (E). Scale bars, 20 μm. (B) Immunostaining for histone H3R17 methylation (green) was performed with DNA counterstained with Hoechst 33342 (blue) and corresponding MFI of quantitative data (F). Scale bars, 20 μm. (C) Immunostaining for histone H3R17 citrullination (green) was performed with DNA counterstained with Hoechst 33342 (blue) and corresponding MFI of quantitative data (G). Scale bars, 20 μm. (D) Histone H3R17 citrullination was regulated by H3R17 methylation inhibitor (EZM2302) and HDAC2 inhibitor (SAHA), and immunostaining for histone H3R17 citrullination (green) was performed with DNA counterstained with Hoechst 33342 (blue), the corresponding MFI of quantitative data (H). Scale bars, 20 μm. (I) H3K18 acetylation, H3R17 methylation and H3R17 citrullination were detected by western blotting in the neutrophils from HDAC2 WT and HDAC2 KO mice after stimulation with LPS (50 μg/ml). The H3 protein was used for western blot loading controls. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Then we cultured them using a secondary Alexa Fluor 488 goat anti-rabbit IgG (abcam, ab150077, 1 μg/ml) with parallel Hoechst 33342 staining (Beyotime Biotech, C1026, 10 μg/ml).

Techniques: Immunostaining, Methylation, Western Blot

Journal: Drug Design, Development and Therapy

Article Title: Atorvastatin Attenuates Vancomycin-Induced Nephrotoxicity via PPARα-Associated Regulation of SLC Transporters

doi: 10.2147/DDDT.S571916

Figure Lengend Snippet: ATO attenuates VCM-induced apoptosis in HK-2 cells. ( A ) Representative images of HK-2 cell apoptosis detected by Hoechst 33342 staining (original magnification ×100); ( B ) Quantification of apoptotic cell percentage; ( C ) Western blot analysis of Bcl-2 and Bax protein expression in HK-2 cells; ( D ) Quantification of Bcl-2 protein levels; ( E ) Quantification of Bax protein levels. The data are presented as mean ± SD, n = 3, analyzed by one-way ANOVA followed by Dunnett’s post hoc test. Compared with the CONTROL group, *** P < 0.001; compared with the VCM model group, # P < 0.05, ## P < 0.01, ### P < 0.001. For comparisons with the VCM model group: ( B ) VCM vs. VCM + ATO (2 μM), P = 0.0002; VCM vs. VCM + ATO (10 μM), P < 0.0001. ( D ) VCM vs. VCM + ATO (2 μM), P = 0.0436; VCM vs. VCM + ATO (10 μM), P = 0.0023. ( E ) VCM vs. VCM + ATO (2 μM), P = 0.0334; VCM vs. VCM + ATO (10 μM), P = 0.0012.

Article Snippet: The Reactive Oxygen Species (ROS) Detection Kit, Hoechst 33342 Staining Solution, Hematoxylin-Eosin (HE) Staining Kit, and TUNEL Kit were purchased from Shanghai Beyotime Biotechnology Co., Ltd. (Shanghai, China).

Techniques: Staining, Western Blot, Expressing, Control