Journal: Neuropsychiatric Disease and Treatment

Article Title: Fibroblast Growth Factor 21 Protects Against Cerebral Ischemia/Reperfusion Injury by Inhibiting Oxidative Stress and Ferroptosis

doi: 10.2147/NDT.S504180

Figure Lengend Snippet: CYBB may be a key effector of FGF21, crucial for inhibiting ferroptosis. ( A ) Venn diagram: Overlap between DEGs and ferroptosis-related genes. ( B ) Comparison of significant differential expression rates for ferroptosis-related vs other genes. ( C ) RNA sequencing: Expression fold changes for the top four significantly altered ferroptosis-related genes. ( D ) qPCR: Relative expression levels for the top four ferroptosis-related genes with significant fold changes. ( E ) Western blotting: CYBB protein expression levels. ( F ) Immunofluorescence double-labeling staining: CYBB and the neuronal-specific marker NeuN. Data represent Mean ± SD; (n = 5). ** P < 0.01, *** P < 0.001. The red arrows: Double-labeled immunofluorescent positive cells.

Article Snippet: The primary antibodies were diluted with 2% BSA at a 1:100 ratio using CYBB (1:100, rabbit, bs-38R, Bioss, Beijing, China) and NeuN (1:100, rabbit, bs-10394R, Bioss).

Techniques: Comparison, Expressing, RNA Sequencing Assay, Western Blot, Immunofluorescence, Labeling, Staining, Marker

Journal: MRS bulletin

Article Title: Functional imaging of brain organoids using high-density microelectrode arrays.

doi: 10.1557/s43577-022-00282-w

Figure Lengend Snippet: Figure 2. Human cerebral organoids comprise diverse cell types relevant to the study of neuronal circuit development and physiology. (a) Immunohistochemistry (IHC) revealed the presence of mature neurons (Tau/NeuN), astrocytes (GFAP), and oligodendrocytes (OLIG2) after 100 days of differentiation; nuclei were stained with DAPI (in blue). Scale bars are 50 μm. (b) Quantification of NeuN-positive cells. The ratio of the number of NeuN-positive cells to the total number of cells was quantified for both hCO lines (3 hCOs per line, for each hCO > 5000 cells). The ratio of NeuN-positive cells to total cells did not differ significantly between the two lines. (c) Single-cell RNA sequencing (scRNA- seq) was performed to quantify the cellular composition of hCOs and their comparability. The results are summarized in two pie charts with the identified cell type fractions displayed for each line. For both lines the results confirmed the presence of dopaminergic, GABAergic, glutamatergic and cholinergic neurons, neural crest cells (NCCs), radial glia (RG), astrocytes and oligodendrocytes.

Article Snippet: The following primary and secondary antibodies were used in this study: Pax6 (BioLegend, San Diego, CA, USA, #901301, 1:300), Sox2 (Sigma-Aldrich, AB5603, 1:300), FoxG1 (Abcam, ab18259, 1:200), Tuj1 (BioLegend, #801202, 1:800), Ctip2 (Abcam, ab18465, 1:200), Tbr1 (Abcam, ab31940, 1:300), MAP2 (Thermo Scientific, PA1-10,005, 1:800), Tau (Thermo Scientific, MN1000, 1:500), NeuN (Boster Bio, Pleasanton, CA, USA, M11954-3, 1:300), GFAP (Novus Biologicals, Englewood, CO, USA, NB300-141 , 1:500), goat anti-mouse IgG (H + L) highly cross-adsorbed secondary antibody, Alexa Fluor Plus 488 (Thermo Scientific, A32723, 1:400), goat anti-rabbit IgG (H + L) highly cross-adsorbed secondary Antibody, Alexa Fluor 568 (Thermo Scientific, A11036, 1:400), goat anti-rat IgG (H + L) cross-adsorbed secondary antibody, Alexa Fluor 647 (Thermo Scientific, A21247, 1:400) and goat anti-chicken IgY (H + L) cross-adsorbed secondary antibody, Alexa Fluor Plus 647 (Thermo Scientific, A32933, 1:400).

Techniques: Immunohistochemistry, Staining, RNA Sequencing

Journal: Mediators of Inflammation

Article Title: Protein Kinase A Is Involved in Neuropathic Pain by Activating the p38MAPK Pathway to Mediate Spinal Cord Cell Apoptosis

doi: 10.1155/2020/6420425

Figure Lengend Snippet: SNI causes neuropathic pain and increases the expression of PKA and p-p38 in the spinal cord. (a, b) SNI ipsilateral PWT (a) and contralateral PWT (b) ( n = 9). ∗∗∗ P < 0.001 versus Sham group. (c–e) Western blots and quantification of PKA and p-p38 expression in the spinal cord ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 versus Sham group. (f) Immunofluorescence images showing PKA expression (green) in the dorsal horn of the spinal cord. (g) Immunofluorescence images showing the distribution of PKA in the cells of the dorsal horn of the spinal cord. PKA is green, NeuN, GFAP, and Iba-1 are red.

Article Snippet: The sections were blocked in 10% goat serum at room temperature for 2 h and incubated at 4°C overnight with primary antibodies against the following proteins: PKA (anti-mouse, 1 : 100, Santa Cruz, Dallas, TX, USA), neuronal nuclei (NeuN) (anti-rabbit, 1 : 50, Proteintech, Wuhan, China), glial fibrillary acidic protein (GFAP) (anti-rabbit, 1 : 500, Servicebio, Wuhan, China), and ionized calcium binding adapter molecule 1 (Iba1) (anti-rabbit, 1 : 500, Servicebio, Wuhan, China).

Techniques: Expressing, Western Blot, Immunofluorescence

Journal: Mediators of Inflammation

Article Title: Protein Kinase A Is Involved in Neuropathic Pain by Activating the p38MAPK Pathway to Mediate Spinal Cord Cell Apoptosis

doi: 10.1155/2020/6420425

Figure Lengend Snippet: PKA and p38MAPK inhibitors reduce inflammation and apoptosis. (a–c) Western blots and quantification of TNF- α and IL-1 β expression in the spinal cord ( n = 3). ∗ P < 0.05 versus Sham group, # P < 0.05, ## P < 0.01 versus SNI group. (d, e) TNF- α (d) and IL-1 β € levels in the spinal cord as measured by ELISA ( n = 3). ∗∗∗ P < 0.001 versus Sham group, ### P < 0.001 versus SNI group. (f–i) Western blots and quantification of bax, bcl-2, caspase3, and caspase9 expression in the spinal cord ( n = 3). ∗ P < 0.05, ∗ P < 0.01 versus Sham group, # P < .05, ## P < 0.01 versus SNI group. (j) TUNEL and NeuN double labeling. TUNEL-positive cells are green; NeuN is red.

Article Snippet: The sections were blocked in 10% goat serum at room temperature for 2 h and incubated at 4°C overnight with primary antibodies against the following proteins: PKA (anti-mouse, 1 : 100, Santa Cruz, Dallas, TX, USA), neuronal nuclei (NeuN) (anti-rabbit, 1 : 50, Proteintech, Wuhan, China), glial fibrillary acidic protein (GFAP) (anti-rabbit, 1 : 500, Servicebio, Wuhan, China), and ionized calcium binding adapter molecule 1 (Iba1) (anti-rabbit, 1 : 500, Servicebio, Wuhan, China).

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Labeling