hoechst 33342 staining solution  (Beyotime)

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: 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

Journal: Foods

Article Title: Protective Effects of Polysaccharides from Pyropia suborbiculata Against UVB-Induced Photodamage in HaCaT Cells

doi: 10.3390/foods15081292

Figure Lengend Snippet: The Effect of PSP-I on UVB-Induced Apoptotic Body Formation. Fluorescent image of Hoechst 33342 observed under a fluorescence microscope. Scale bar = 250 micrometers. White arrows indicate areas of cells exhibiting distinct apoptotic features.

Article Snippet: Hoechst 33342 staining solution was purchased from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China).

Techniques: Fluorescence, Microscopy

Journal: CNS Neuroscience & Therapeutics

Article Title: Forsythoside A Alleviates Neuroinflammatory Damage via Inhibiting TLR4 / NF ‐ κB / NLRP3 Activation‐Induced Astrocyte Pyroptosis in Cerebral Ischemia–Reperfusion

doi: 10.1002/cns.70867

Figure Lengend Snippet: FA suppresses OGD/R astrocyte pyroptosis and inflammation through hindering NLRP3 inflammasome activation. (A–B) Hoechst 33,342 and PI staining showed that 80 μM FA reduced pyroptosis in U251 cells after OGD/R, whereas NLRP3 agonist Nig resulted in increased pyroptosis (20×, 100 μm). (C) LDH assay showed that LDH release from U251 cells was increased following OGD/R FA inhibited LDH release, while Nig led to a rebound in LDH release. (D–E) Immunofluorescence co‐localization measured that OGD/R resulted in increased GSDMD‐N fluorescence intensity in U251 cells, FA treatment decreased GSDMD‐N level, and Nig attenuated the effect of FA (20×, 100 μm). (F–I) ELISA indicated that OGD/R resulted in increased IL‐1β, TNF‐α, IL‐18, and IL‐6 levels; FA treatment declined these inflammatory factors, but addition of Nig resulted in increased levels. (J–M) Western blot measured that OGD/R caused elevated iNOS, IL‐1β, and GSDMD‐N levels in U251 cells, and FA treatment decreased these protein levels, whereas the addition of Nig caused a rebound in these protein levels. (N) SEM to observe the morphological changes associated with pyroptosis in U251 cells. *** p < 0.001 vs. Control; ** p < 0.01 vs. OGD/R; && p < 0.01 vs. 80 μM FA.

Article Snippet: Hoechst 33,342 staining solution (C1025, Beyotime) was introduced and cultured for half an hour at 4°C.

Techniques: Activation Assay, Staining, Lactate Dehydrogenase Assay, Immunofluorescence, Fluorescence, Enzyme-linked Immunosorbent Assay, Western Blot, Control