staining solutions a  (Keygen Biotech)

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: A dual-functional hydrogel integrating adhesive and lubricating interfaces for mitochondrial protection–Driven cartilage regeneration

doi: 10.1016/j.bioactmat.2026.02.051

Figure Lengend Snippet: In vitro antioxidant and mitochondrial homeostasis regulatory effects of AdHy@Pae. (A) Flow cytometric analysis of the cell cycle. (B–D) Quantitative statistics of the percentage of cells in G0/G1, S, and G2/M phases. (E) JC-1 fluorescence staining. (F) Quantitative analysis of JC-1 red/green fluorescence ratio (ΔΨm). (G) Flow cytometric detection of intracellular ROS using DCFH-DA probe. Data are shown as mean ± SD (n = 3, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001).

Article Snippet: After LPS stimulation (10 μg mL −1 , 12 h, ServiceBio, GC205009 ) to induce an inflammatory phenotype, cells were treated with hydrogel extracts (Hy, AdHy, AdHy@Pae) for 24 h. Cells were then washed twice with PBS, harvested, and fixed overnight at 4 °C in 70% cold ethanol; after washing, they were stained with 500 μL cell-cycle staining solution (MULTI SCIENCES, CCS012) for 30 min in the dark.

Techniques: In Vitro, Fluorescence, Staining

Journal: Genes & Diseases

Article Title: Non-canonical role of “S6K1–SGK1” pathway in neuronal necroptosis following traumatic brain injury

doi: 10.1016/j.gendis.2025.101876

Figure Lengend Snippet: S6K1 inhibition alleviates neuronal necroptosis through SGK1 following TBI in mice. (A, K) Western blotting showed increased expression of p-S6/S6, SGK1, p-MLKL/MLKL, and p-RIP3/RIP3 in the cortex following TBI. These changes were attenuated by S6K1 inhibitor treatment and S6K1 knockdown, as shown by decreased p-MLKL/MLKL and p-RIP3/RIP3 levels compared with the TBI group. (B–E, L, M) Statistical results of Western blotting. (F) Immunohistochemistry staining of p-S6K1 in the right cortex of mouse brain after TBI. Scale bar: 20 μm. (G) Statistical results of immunohistochemistry staining for the increased S6K1 expression. (H) Schematic illustration of protein detection and behavior paradigm following p-S6K1 inhibitor injection. (I) Nissl staining of decreased neurons after TBI, and increased neurons after treatment with S6K1 inhibitor and knockdown compared with TBI. Scale bar: 20 μm. (J) Statistical results of Nissl staining for the numbers of neurons. (N) Enhanced immunofluorescence intensity of p-MLKL in the right cortex of mouse brain after TBI, and decreased immunofluorescence intensity after treatment with S6K1 inhibitor and knockdown compared with TBI. Scale bar: 50 μm. Two-tailed unpaired Student's t -test was used for (G). n = 3 or 4. ∗ P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001 versus the Sham group. # P < 0.05 and ## P < 0.01 versus the TBI group.

Article Snippet: The brain sections were dewaxed and stained with Nissl staining solution (Beyotime) for 10 min.

Techniques: Inhibition, Western Blot, Expressing, Knockdown, Immunohistochemistry, Staining, Injection, Immunofluorescence, Two Tailed Test