hoechst 33342  (Miltenyi Biotec)

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