primary antibodies against ox1r  (Proteintech)

Journal: CNS Neuroscience & Therapeutics

Article Title: Neuroprotective Effects of Transcranial Pulsed Current Stimulation: Modulation of Microglial Polarization in Traumatic Brain Injury

doi: 10.1111/cns.70606

Figure Lengend Snippet: The impact of tPCS on the expression of OX‐A, OX1R, MAP2, GAP43, and TGF‐β in the peri‐lesional cortex of TBI mice. (A, B) Transcriptome sequencing identified DEGs between the TBI and tPCS groups, as visualized in a volcano plot, and KEGG enrichment analysis was performed on these DEGs ( n = 3). (C–G) Representative Western blot images and quantification illustrate the levels of OX‐A, OX1R, GAP43, and MAP2 in the peri‐lesional cortex at 7 dpo ( n = 4), with β‐actin serving as the loading control. The original blots are presented in the Appendix . (H–M) Representative IF images and quantification depict the expression levels of OX1R, GAP43, and MAP2 at 7 dpo ( n = 4). (N–P) Representative IF images and quantitative analysis of NeuN and TGF‐β expression levels in the peri‐lesional cortex ( n = 4). Scale bar =100 μm. All values are mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Subsequently, primary antibodies against OX1R (183701AP, Proteintech, China), ZO‐1 (ab307799, Abcam, USA), MAP2 (ab254264, Abcam), GAP43 (ab277627, Abcam), TGF‐β1 (ab215715, Abcam), IBA1 (ab178846, Abcam), Bcl‐xL (ab270253, Abcam), NeuN (ab104224, Abcam), CD31 (ab182981, Abcam), BrdU (Ab6326, Abcam), CD206 (#91992, Cell Signaling Technology), and CD86 (ab119857, Abcam) were used to the sections and incubated at 4°C for 12 h. The sections were then incubated with Alexa Fluor 488 or 555‐conjugated secondary antibodies for 2 h. Fluorescence microscopy was used to capture images from three randomly selected, nonoverlapping ROIs in the cortex.

Techniques: Expressing, Sequencing, Western Blot, Control

Journal: CNS Neuroscience & Therapeutics

Article Title: Neuroprotective Effects of Transcranial Pulsed Current Stimulation: Modulation of Microglial Polarization in Traumatic Brain Injury

doi: 10.1111/cns.70606

Figure Lengend Snippet: tPCS attenuated neuronal apoptosis and ameliorated neurological deficits via OX‐A/OX1R in TBI mice. (A–E) Motor function of TBI mice was assessed using Y‐maze, beam‐balance test, and rotarod test ( n = 8). (F–H) Representative IF images and quantitative analysis of IBA1 and OX1R expression levels in the peri‐lesional cortex ( n = 6). The arrows represent OX1R+IBA1‐positive cells. (I–K) Representative IF images and quantitative analysis of Bcl‐xL and NeuN ( n = 6). (L) Representative images of the cortical region were obtained using HE staining, Nissl staining, and transmission electron microscopy. (M) Quantification of Nissl‐stained cells ( n = 6). (N) Representative images of the cortical region were obtained using transmission electron microscopy. (O–Q) Representative Western blot bands and quantitative analysis demonstrating Bcl‐xL and NeuN levels in the peri‐lesional cortex ( n = 5), with β‐actin employed as the loading control. Scale bar = 100 μm. The original blots are presented in Appendix . All values are mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Subsequently, primary antibodies against OX1R (183701AP, Proteintech, China), ZO‐1 (ab307799, Abcam, USA), MAP2 (ab254264, Abcam), GAP43 (ab277627, Abcam), TGF‐β1 (ab215715, Abcam), IBA1 (ab178846, Abcam), Bcl‐xL (ab270253, Abcam), NeuN (ab104224, Abcam), CD31 (ab182981, Abcam), BrdU (Ab6326, Abcam), CD206 (#91992, Cell Signaling Technology), and CD86 (ab119857, Abcam) were used to the sections and incubated at 4°C for 12 h. The sections were then incubated with Alexa Fluor 488 or 555‐conjugated secondary antibodies for 2 h. Fluorescence microscopy was used to capture images from three randomly selected, nonoverlapping ROIs in the cortex.

Techniques: Expressing, Staining, Transmission Assay, Electron Microscopy, Western Blot, Control

Journal: CNS Neuroscience & Therapeutics

Article Title: Neuroprotective Effects of Transcranial Pulsed Current Stimulation: Modulation of Microglial Polarization in Traumatic Brain Injury

doi: 10.1111/cns.70606

Figure Lengend Snippet: tPCS enhances the expression of neurorepair‐related markers and neuroplasticity following TBI via the OX‐A/OX1R pathway. (A, B) Representative IF images and quantification of NeuN+BrdU‐positive cells in the peri‐lesional cortex ( n = 6). The arrows represent NeuN+BrdU‐positive cells. Scale bar = 100 μm. (C, D) Representative IF images and quantification of CD31+BrdU‐positive cells ( n = 6). The arrows represent CD31+BrdU positive cells. Scale bar = 100 μm. (E) Quantification of the brain water content ( n = 3). (F–H) Representative IF images and quantitative analysis of ZO‐1 and CD31 in the peri‐lesional cortex ( n = 6). Scale bar = 100 μm. (I, J) Representative images and quantitative analysis of Evans Blue staining at 7dpo ( n = 4). (K, L) Representative images of Golgi staining and Sholl analysis in the peri‐lesional cortex ( n = 6). The red dotted box represents the neurons selected for Sholl analysis. Scale bar =50 μm. (M, N) Representative imaging of dendrites and quantification of dendritic density ( n = 6). (O–Q) SYP and PSD95 proteins were measured and quantified at 7 dpo by Western blot ( n = 5), with β‐actin serving as a loading control. The original blots are presented in Appendix . (R, S) Representative TEM images and quantification of postsynaptic densities in the perilesional cortex. Scale bar = 1 μm. All values are mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Subsequently, primary antibodies against OX1R (183701AP, Proteintech, China), ZO‐1 (ab307799, Abcam, USA), MAP2 (ab254264, Abcam), GAP43 (ab277627, Abcam), TGF‐β1 (ab215715, Abcam), IBA1 (ab178846, Abcam), Bcl‐xL (ab270253, Abcam), NeuN (ab104224, Abcam), CD31 (ab182981, Abcam), BrdU (Ab6326, Abcam), CD206 (#91992, Cell Signaling Technology), and CD86 (ab119857, Abcam) were used to the sections and incubated at 4°C for 12 h. The sections were then incubated with Alexa Fluor 488 or 555‐conjugated secondary antibodies for 2 h. Fluorescence microscopy was used to capture images from three randomly selected, nonoverlapping ROIs in the cortex.

Techniques: Expressing, Staining, Imaging, Western Blot, Control

Journal: CNS Neuroscience & Therapeutics

Article Title: Neuroprotective Effects of Transcranial Pulsed Current Stimulation: Modulation of Microglial Polarization in Traumatic Brain Injury

doi: 10.1111/cns.70606

Figure Lengend Snippet: Treatment with tPCS modulates M1/M2 microglia polarization via OX‐A/OX1R‐mediated NF‐κB pathway following TBI. (A, B) Representative IF images and quantification of CD86+IBA1‐positive cells in the peri‐lesional cortex ( n = 6). (C, D) Representative IF image and quantification of CD206+IBA1‐positive cells ( n = 6). (E–G) Representative immunofluorescence image and quantification of p65+IBA1‐positive cells ( n = 6). (H, I) Representative IF image and quantitative analysis of NeuN and TGF‐β ( n = 6). (J–N) Representative Western blot bands and quantification showing levels of phosphorylated (p)‐p65, p‐IκBα, iNOS, and Arg‐1 proteins in the peri‐lesional cortex ( n = 5), with β‐actin serving as a loading control. All values are mean ± SD. Scale bar = 100 μm. The original blots are presented in Appendix . * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Subsequently, primary antibodies against OX1R (183701AP, Proteintech, China), ZO‐1 (ab307799, Abcam, USA), MAP2 (ab254264, Abcam), GAP43 (ab277627, Abcam), TGF‐β1 (ab215715, Abcam), IBA1 (ab178846, Abcam), Bcl‐xL (ab270253, Abcam), NeuN (ab104224, Abcam), CD31 (ab182981, Abcam), BrdU (Ab6326, Abcam), CD206 (#91992, Cell Signaling Technology), and CD86 (ab119857, Abcam) were used to the sections and incubated at 4°C for 12 h. The sections were then incubated with Alexa Fluor 488 or 555‐conjugated secondary antibodies for 2 h. Fluorescence microscopy was used to capture images from three randomly selected, nonoverlapping ROIs in the cortex.

Techniques: Immunofluorescence, Western Blot, Control