Journal: Neural Regeneration Research

Article Title: High-dose dexamethasone regulates microglial polarization via the GR/JAK1/STAT3 signaling pathway after traumatic brain injury

doi: 10.4103/NRR.NRR-D-23-01772

Figure Lengend Snippet: Microglial activation in response to LPS and DEX treatment. (A) Representative immunofluorescence images showing that DEX treatment decreases M1 and M2 microglial polarization in cultured BV2 cells and primary microglia after exposure to LPS. Scale bars: 20 μm. Red represents Iba1, and green represents Arg1 or iNOS. (B) Counts of Arg1- and iNOS-positive cells normalized to Iba1-positive cells. * P < 0.05, vs . Control; # P < 0.05, vs . LPS (one-way analysis of variance with the least significant difference test). Data are presented as mean ± SD, n = 3. DAPI: 4′,6-Diamidino-2-phenylindole; DEX: dexamethasone; LPS: lipopolysaccharide.

Article Snippet: After blocking, cells were incubated with primary antibodies: rabbit monoclonal anti-ionized calcium binding adaptor molecule 1 (Iba1) (1:200, Abcam, Cat# ab289370), mouse monoclonal anti-Arg1 (1:100, Abcam, Cat# ab239731), and mouse monoclonal anti-inducible nitric oxide synthase (iNOS) (1:500, Abcam, Cat# ab210823) for 24 hours at 4°C.

Techniques: Activation Assay, Immunofluorescence, Cell Culture, Control

Journal: Neural Regeneration Research

Article Title: High-dose dexamethasone regulates microglial polarization via the GR/JAK1/STAT3 signaling pathway after traumatic brain injury

doi: 10.4103/NRR.NRR-D-23-01772

Figure Lengend Snippet: Distribution of DEGs between TBI and DEX groups. (A) Heatmap displaying hierarchical clustering of DEGs in the cortex. (B) Hierarchical clustering of DEGs in the hippocampus. Blue indicates low expression levels, and red indicates high expression levels. TBI-C: Cortex of the TBI group; TBI-H: Hippocampus of the TBI group; DEX-C: Cortex of the dexamethasone (DEX) group; DEX-H: Hippocampus of the DEX group. (C) Volcano plot showing DEGs in the cortex. (D) Expression of DEGs in the hippocampus. Upregulated and downregulated DEGs are highlighted in red and blue, respectively. (E) Gene sequencing map showing expression levels of Arg1 in the cortex. (F) Expression levels of Arg1 in the hippocampus. Plot color intensity and size separately indicate degree of the P -value and log2(fold change) value. DEGs were determined based on absolute log2(fold change) > 1 and P < 0.05. n = 3 individuals each in TBI and DEX groups. DEGs: Differentially expressed genes; DEX-C: cortex of dexamethasone; DEX-H: hippocampus of dexamethasone; TBI: traumatic brain injury; TBI-C: cortex of traumatic brain injury; TBI-H: hippocampus of traumatic brain injury.

Article Snippet: After blocking, cells were incubated with primary antibodies: rabbit monoclonal anti-ionized calcium binding adaptor molecule 1 (Iba1) (1:200, Abcam, Cat# ab289370), mouse monoclonal anti-Arg1 (1:100, Abcam, Cat# ab239731), and mouse monoclonal anti-inducible nitric oxide synthase (iNOS) (1:500, Abcam, Cat# ab210823) for 24 hours at 4°C.

Techniques: Expressing, Sequencing

Journal: Neural Regeneration Research

Article Title: High-dose dexamethasone regulates microglial polarization via the GR/JAK1/STAT3 signaling pathway after traumatic brain injury

doi: 10.4103/NRR.NRR-D-23-01772

Figure Lengend Snippet: Microglial activation in response to LPS and DEX treatment. (A) Representative immunofluorescence images showing that DEX treatment decreases M1 and M2 microglial polarization in cultured BV2 cells and primary microglia after exposure to LPS. Scale bars: 20 μm. Red represents Iba1, and green represents Arg1 or iNOS. (B) Counts of Arg1- and iNOS-positive cells normalized to Iba1-positive cells. * P < 0.05, vs . Control; # P < 0.05, vs . LPS (one-way analysis of variance with the least significant difference test). Data are presented as mean ± SD, n = 3. DAPI: 4′,6-Diamidino-2-phenylindole; DEX: dexamethasone; LPS: lipopolysaccharide.

Article Snippet: After blocking, cells were incubated with primary antibodies: rabbit monoclonal anti-ionized calcium binding adaptor molecule 1 (Iba1) (1:200, Abcam, Cat# ab289370), mouse monoclonal anti-Arg1 (1:100, Abcam, Cat# ab239731), and mouse monoclonal anti-inducible nitric oxide synthase (iNOS) (1:500, Abcam, Cat# ab210823) for 24 hours at 4°C.

Techniques: Activation Assay, Immunofluorescence, Cell Culture, Control

Journal: Neural Regeneration Research

Article Title: High-dose dexamethasone regulates microglial polarization via the GR/JAK1/STAT3 signaling pathway after traumatic brain injury

doi: 10.4103/NRR.NRR-D-23-01772

Figure Lengend Snippet: Reduction in M2 microglial expression by DEX is regulated by GR/JAK1/STAT3 pathway activation. (A) qRT-PCR detected M2 markers, Arg1, JAK1, STAT3, Bcl2, and Myc. (B) Western blotting measured activation levels of glucocorticoid receptors (GR) (83 kDa), and expression levels of Arg1 (35 kDa), JAK1 (133 kDa), p-STAT3 (98 kDa), Bcl2 (26 kDa), c-Myc (57 kDa) and GAPDH (36 kDa). Data are expressed as mean ± SD, n = 3. * P < 0.05, vs . Control; # P < 0.05, vs. TBI (one-way analysis of variance with the least significant difference test). DEX: Dexamethasone; TBI: traumatic brain injury.

Article Snippet: After blocking, cells were incubated with primary antibodies: rabbit monoclonal anti-ionized calcium binding adaptor molecule 1 (Iba1) (1:200, Abcam, Cat# ab289370), mouse monoclonal anti-Arg1 (1:100, Abcam, Cat# ab239731), and mouse monoclonal anti-inducible nitric oxide synthase (iNOS) (1:500, Abcam, Cat# ab210823) for 24 hours at 4°C.

Techniques: Expressing, Activation Assay, Quantitative RT-PCR, Western Blot, Control

Journal: Neural Regeneration Research

Article Title: High-dose dexamethasone regulates microglial polarization via the GR/JAK1/STAT3 signaling pathway after traumatic brain injury

doi: 10.4103/NRR.NRR-D-23-01772

Figure Lengend Snippet: Expression patterns of Arg1 following TBI. (A, B) Representative immunofluorescence staining (A) and quantification (B) showing expression levels of Arg1 in the peri-injured cortex and hippocampus after TBI. Inhibition of glucocorticoid receptor activation reversed M2 microglial marker expression. Red represents Arg1, and blue represents DAPI. Scale bars: 20 μm. Data are expressed as mean ± SD, n = 3. * P < 0.05, vs . Control; # P < 0.05, vs. TBI (one-way analysis of variance with the least significant difference test). DAPI: 4′,6-Diamidino-2-phenylindole; DEX: dexamethasone; TBI: traumatic brain injury.

Article Snippet: After blocking, cells were incubated with primary antibodies: rabbit monoclonal anti-ionized calcium binding adaptor molecule 1 (Iba1) (1:200, Abcam, Cat# ab289370), mouse monoclonal anti-Arg1 (1:100, Abcam, Cat# ab239731), and mouse monoclonal anti-inducible nitric oxide synthase (iNOS) (1:500, Abcam, Cat# ab210823) for 24 hours at 4°C.

Techniques: Expressing, Immunofluorescence, Staining, Inhibition, Activation Assay, Marker, Control

Journal: Neural Regeneration Research

Article Title: Small molecule inhibitor DDQ-treated hippocampal neuronal cells show improved neurite outgrowth and synaptic branching

doi: 10.4103/NRR.NRR-D-24-00157

Figure Lengend Snippet: Antibodies used in western blot analysis

Article Snippet: MAP2 , Mouse monoclonal 1:500 , 13–1500 , Invitrogen, Carlsbad, CA, USA , , Anti-mouse IgG 1:1000 , NA931V , Invitrogen.

Techniques: Western Blot

Journal: Neural Regeneration Research

Article Title: Small molecule inhibitor DDQ-treated hippocampal neuronal cells show improved neurite outgrowth and synaptic branching

doi: 10.4103/NRR.NRR-D-24-00157

Figure Lengend Snippet: Antibodies used in immunofluorescence analysis

Article Snippet: MAP2 , Mouse monoclonal 1:500 , 13–1500 , Invitrogen, Carlsbad, CA, USA , , Anti-mouse IgG 1:1000 , NA931V , Invitrogen.

Techniques: Immunofluorescence