pc12 neuronal cell lines  (ATCC)

Journal: Neural Regeneration Research

Article Title: Inhibiting SHP2 reduces glycolysis, promotes microglial M1 polarization, and alleviates secondary inflammation following spinal cord injury in a mouse model

doi: 10.4103/NRR.NRR-D-23-01925

Figure Lengend Snippet: SHP2 inhibition reduces neuronal apoptosis in the context of SCI. (A, B) NeuN (red-CoraLite594) and SHP2 (green-CoraLite488) immunoreactivities in mice treated with NSC87877 14 days after SCI, as detected by immunofluorescence. Quantitative analysis showing that a decrease in SHP2 fluorescence intensity and an increase in the fluorescence intensity of NeuN, a neuronal marker, in SCI + NSC87877 group mice near the injury site ( n = 5 slices from three mice per group). Scale bars: 50 μm. (C) Nissl staining of mouse spinal cord transverse and longitudinal sections 14 days after SCI showing that, compared with the SCI group, the SCI + NSC87877 group exhibited more Nissl bodies with a more uniform distribution ( n = 5 slices from three mice per group). Scale bars: 100 μm (left), 600 μm (right). (D, E) Western blot analysis of Caspase3, C-caspase3, Bax, and Bcl-2 protein expression in PC12 cells co-cultured with BV2 cells treated with LPS or LPS + NSC87877 for 24 hours. The experiment was repeated three times. (F, G) C-caspase3 (red-CoraLite594) expression in PC12 cells was detected by immunofluorescence staining. Quantitative analysis showed decreased immunofluorescence intensity of the apoptosis marker C-caspase3 in neurons co-cultured with BV2 cells from the LPS + NSC87877 group. The experiment was repeated three times. Scale bars: 50 μm. Data are expressed as the mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance with Bonferroni’s post hoc test). Bax: Bcl-2-associated X protein; Bcl-2: B-cell lymphoma 2; Caspase3: cysteine-aspartic acid protease 3; C-caspase3: cleaved caspase-3; DAPI: 4′,6-diamidino-2′-phenylindole; IF: immunofluorescence staining; LPS: lipopolysaccharide; NeuN: neuronal nuclei; NSC87877: SHP2 inhibitor; SCI: spinal cord injury; SHP2: Src homology 2 domain-containing protein tyrosine phosphatase 2.

Article Snippet: The PC12 neuronal cell line (Cat# CL-0481, RRID:CVCL_0481), and the BV2 mouse microglia cell line (Cat# CL0493, RRID: CVCL_0182) were purchased from Procell Life Science and Technology Co. (Wuhan, China).

Techniques: Inhibition, Immunofluorescence, Fluorescence, Marker, Staining, Western Blot, Expressing, Cell Culture

Journal: CNS Neuroscience & Therapeutics

Article Title: Activation of STING signaling aggravates chronic alcohol exposure‐induced cognitive impairment by increasing neuroinflammation and mitochondrial apoptosis

doi: 10.1111/cns.14689

Figure Lengend Snippet: Activation of STING signaling induced by alcohol in neurons led to mitochondrial apoptosis. (A, B) Representative immunoblot images and quantitative analyses of the expression of STING pathway‐associated genes in PC12 cells treated with control and EtOH (300 mM) ( n = 3). (C) Representative immunoblot images showing the expression of STING signaling pathway‐associated genes in primary neurons treated with various concentrations of alcohol (0, 100, 200, 300 mM). (D, E) Representative images and quantitative analyses of apoptotic PC12 cells in the different groups, as measured by flow cytometry ( n = 4, DMXAA (50 μg/mL)). (F) Quantitative analyses of the activities of caspase 3, caspase 9, and caspase 8 in PC12 cells in the different groups ( n = 3, DMXAA (50 μg/mL)). (G, I) Effects of DMXAA on the mitochondrial membrane potential of PC12 cells treated with alcohol, as measured by flow cytometry using JC‐1 ( n = 3, DMXAA (50 μg/mL)). (H, J) Effects of DMXAA on ROS production in PC12 cells treated with alcohol, as measured by flow cytometry using JC‐1 ( n = 3, DMXAA (50 μg/mL)). Scale bar = 100 μm. (K, L) Representative images and quantitative analyses of TBK1, p‐TBK1 and mitochondrial apoptosis‐associated gene expression in the different groups ( n = 3, DMXAA (50 μg/mL)). The data are expressed as the mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001; ns, nonsignificant.

Article Snippet: The mouse microglial cell line BV2 and the mouse neuronal cell line PC12, purchased from Procell Life, China, were cultured in DMEM (Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Excellbio, China) and 1% penicillin and streptomycin (Gibco, USA) at 37°C and 5% CO2.

Techniques: Activation Assay, Western Blot, Expressing, Control, Flow Cytometry, Membrane, Gene Expression