Journal: Neural Regeneration Research

Article Title: Dynamic development of microglia and macrophages after spinal cord injury

doi: 10.4103/NRR.NRR-D-24-00063

Figure Lengend Snippet: qRT-PCR primers

Article Snippet: After blocking with 5% non-fat milk for 1 hour, membranes were incubated overnight at 4°C with primary antibodies rabbit anti-ITGB2 (1:1000; Proteintech; Cat# 10554-1-AP, RRID: AB_2877736) and rabbit anti-β-tubulin (1:30,000; Proteintech; Cat# 66240-1, RRID: AB_2881629).

Techniques:

Journal: Neural Regeneration Research

Article Title: Dynamic development of microglia and macrophages after spinal cord injury

doi: 10.4103/NRR.NRR-D-24-00063

Figure Lengend Snippet: Expression of Itgb2 in different cell types in SCI model mice. (A) t-SNE plot of the annotated gene Itgb2 in SCI model mice. (B) Expression levels of Itgb2 in different cell lineages in the sham, 1 dpi, 3 dpi, and 7 dpi groups. (C) SpaTalk cell-type decomposition at single-cell resolution for the spot-based spatial transcriptomics data at 7 dpi. (D) Visium spot-based spatial transcriptomics dataset of the SCI mouse model and the SpaTalk cell-type decomposition showing the percentage of Itgb2 expression level. (E, F) Spatial distribution of microglia (E) and macrophages (F). DCs: Dendritic cells; dpi: Day(s) post injury; Itgb2: integrin β2; MAC: macrophages; MONO: monocytes; NEUT: neutrophils; Oligo: oligodendrocytes; T: T cells; SCI: spinal cord injury; ST: spatial transcriptomics; t-SNE: t-distributed stochastic neighbor embedding.

Article Snippet: After blocking with 5% non-fat milk for 1 hour, membranes were incubated overnight at 4°C with primary antibodies rabbit anti-ITGB2 (1:1000; Proteintech; Cat# 10554-1-AP, RRID: AB_2877736) and rabbit anti-β-tubulin (1:30,000; Proteintech; Cat# 66240-1, RRID: AB_2881629).

Techniques: Expressing

Journal: Neural Regeneration Research

Article Title: Dynamic development of microglia and macrophages after spinal cord injury

doi: 10.4103/NRR.NRR-D-24-00063

Figure Lengend Snippet: Validation of Itgb2 expression in spinal cord tissue in vivo . (A) Schematic diagram of experiments used to validate Itgb2 expression in the C57BL/6J SCI mouse model in vivo . (B, C) Western blotting (B) and quantification (C) of ITGB2 protein expression in the sham, 1 dpi, 3 dpi, and 7 dpi groups ( n = 4 for each group). (D) qRT-PCR validation of Itgb2 expression in the sham ( n = 4), 1 dpi ( n = 4), 3 dpi ( n = 4), and 7 dpi ( n = 3) groups. (E) Counting of total number of ITGB2 + cells merged with DAPI-positive cells in F. (F) Representative immunofluorescence (IF) staining of ITGB2 (yellow, Alexa Fluor ®-555), in the sham, 1 dpi, 3 dpi, and 7 dpi groups, showing a rapid increase in the number of ITGB2-positive cells post-SCI that gradually decreased over time. Nuclei were stained with DAPI (blue). The core injury zone is outlined in each image. Scale bar: 500 μm. Areas outlined in red boxes were magnified. * P < 0.05, *** P < 0.001, **** P < 0.0001. Data expressed as mean ± standard deviation ( n = 5 for each group) and analyzed by one-way analysis of variance using Tukey’s multiple comparisons test. DAPI: 4′,6-Diamidino-2-phenylindole; dpi: day(s) post injury; Itgb2: integrin β2; ns: not significant; qRT–PCR: quantitative reverse transcription–polymerase chain reaction; SCI: spinal cord injury.

Article Snippet: After blocking with 5% non-fat milk for 1 hour, membranes were incubated overnight at 4°C with primary antibodies rabbit anti-ITGB2 (1:1000; Proteintech; Cat# 10554-1-AP, RRID: AB_2877736) and rabbit anti-β-tubulin (1:30,000; Proteintech; Cat# 66240-1, RRID: AB_2881629).

Techniques: Biomarker Discovery, Expressing, In Vivo, Western Blot, Quantitative RT-PCR, Immunofluorescence, Staining, Standard Deviation, Reverse Transcription, Polymerase Chain Reaction