Journal: Journal of neuroinflammation

Article Title: Dynamic phosphorylation of Fascin-1 orchestrates microglial phagocytosis and neurological recovery after spinal cord injury.

doi: 10.1186/s12974-025-03445-z

Figure Lengend Snippet: Fig. 9 The function of the balance of Fascin-1 phosphorylation in modulating microglial phagocytosis following SCI. Microglia has extremely robust phagocytic activity in early injury stage (3 dpi), but appears phagocytosis blockage in subacute stage (7 dpi). Pharmacological intervention targeting the Mas1/PKCγ axis with captopril effectively restores the balance of Fascin-1 phosphorylation and microglial phagocytosis

Article Snippet: Briefly, the sections were blocked in PBS buffer containing 10% donkey serum albumin (DSA, SL050, Solarbio, China) and 0.3% Triton X-100 (T8200, Solarbio, China) at room temperature for 1 h. Then, the sections were incubated with primary antibodies at 4 °C overnight, followed by incubation with corresponding secondary antibodies at room temperature for 1 h. The primary antibodies utilized were as follows: mouse antiFascin-1 antibody (1:50, sc-21743, Santa Cruz, United States), rabbit anti-p-Fascin-1 antibody (1:100, ab90618, Abcam, United States), mouse anti-Mas1 antibody (1:50, sc-390453, Santa Cruz, United States), rabbit antiCx3cr1 antibody (1:100, ab8021, Abcam, United States), mouse anti-Cx3cr1 antibody (1:50, sc-377227, Santa Cruz, United States), rat anti-GFAP antibody (1:100, 13–0300, Invitrogen, United States), rabbit anti-NeuN antibody (1:100, 24307, CST, United States), rabbit antiMBP antibody (1:100, AB5864, Merck, Germany), mouse anti-PKCα antibody (1:50, sc-8393, Santa Cruz, United States).

Techniques: Phospho-proteomics, Activity Assay

Journal: Journal of neuroinflammation

Article Title: Dynamic phosphorylation of Fascin-1 orchestrates microglial phagocytosis and neurological recovery after spinal cord injury.

doi: 10.1186/s12974-025-03445-z

Figure Lengend Snippet: Fig. 2 Fascin-1 regulates microglial phagocytosis in vivo and vitro (A) Representative immunofluorescence images of Cx3cr1+ microglia (red) and MBP (green) at 1, 3 and 7 dpi in Fascin-1fl/fl and Fascin-1 CKO mice. White arrowheads indicate microglia phagocytosing MBP. (B) Quantification of the percent age of Cx3cr1+ cells containing MBP in (A). Hollow data points represent values from male mice. Solid data points denote female mice. (C) Representative immunofluorescence images of primary microglia (phalloidin staining, red) incubated with Dio-myelin (1 mg/ml, green) at 0–5 d in vitro. (D) Quantifica tion of intensity of Dio-Myelin staining/cell in (C). Scale bars: 5 μm (A) and 20 μm (C). Data are presented as means ± SD. n = 7 mice per group in (B). n = 3 independent cultures in (D). *p < 0.05, **p < 0.01, ***p < 0.001 by twoway ANOVA followed by Tukey’s post hoc test

Article Snippet: Briefly, the sections were blocked in PBS buffer containing 10% donkey serum albumin (DSA, SL050, Solarbio, China) and 0.3% Triton X-100 (T8200, Solarbio, China) at room temperature for 1 h. Then, the sections were incubated with primary antibodies at 4 °C overnight, followed by incubation with corresponding secondary antibodies at room temperature for 1 h. The primary antibodies utilized were as follows: mouse antiFascin-1 antibody (1:50, sc-21743, Santa Cruz, United States), rabbit anti-p-Fascin-1 antibody (1:100, ab90618, Abcam, United States), mouse anti-Mas1 antibody (1:50, sc-390453, Santa Cruz, United States), rabbit antiCx3cr1 antibody (1:100, ab8021, Abcam, United States), mouse anti-Cx3cr1 antibody (1:50, sc-377227, Santa Cruz, United States), rat anti-GFAP antibody (1:100, 13–0300, Invitrogen, United States), rabbit anti-NeuN antibody (1:100, 24307, CST, United States), rabbit antiMBP antibody (1:100, AB5864, Merck, Germany), mouse anti-PKCα antibody (1:50, sc-8393, Santa Cruz, United States).

Techniques: In Vivo, Immunofluorescence, Staining, Incubation, In Vitro

Journal: Journal of neuroinflammation

Article Title: Dynamic phosphorylation of Fascin-1 orchestrates microglial phagocytosis and neurological recovery after spinal cord injury.

doi: 10.1186/s12974-025-03445-z

Figure Lengend Snippet: Fig. 8 Intraperitoneal injection of captopril protected residual neurons and promotes locomotor recovery after SCI (A) Mice were injected intraperitone ally with captopril (39 mg/kg) or the control solvent immediately after surgery until 28 dpi. (B) Representative immunofluorescence images of GFAP+ astrocytes (green) and NeuN+ neurons (red) in Z1-Z4 zones adjacent to the lesion core in mice treated with or without captopril (39 mg/kg) at 28 dpi. R and C represent the rostral and caudal sides of the injury respectively. (C) Quantification of NeuN+ neurons numbers in (B). (D) Behavioral testing using the BMS score was performed at 0–28 dpi in mice treated with or without captopril (39 mg/kg). *p < 0.05, **p < 0.01, ***p < 0.001 in (C) and (D) by twoway ANOVA followed by Tukey’s post hoc test. (E) Footprint assays in mice treated with or without captopril (39 mg/kg) at 28 dpi. (F-H) Quantification of Paw rotation (F) Stride length (G) Stride width (H) in Footprint assays. Hollow data points represent values from male mice. Solid data points denote female mice. Scale bars: 100 μm (B). Data are presented as means ± SD. n = 7 mice per group in (C-H). *p < 0.05, **p < 0.01, ***p < 0.001 in (F-H) by Student’s t test

Article Snippet: Briefly, the sections were blocked in PBS buffer containing 10% donkey serum albumin (DSA, SL050, Solarbio, China) and 0.3% Triton X-100 (T8200, Solarbio, China) at room temperature for 1 h. Then, the sections were incubated with primary antibodies at 4 °C overnight, followed by incubation with corresponding secondary antibodies at room temperature for 1 h. The primary antibodies utilized were as follows: mouse antiFascin-1 antibody (1:50, sc-21743, Santa Cruz, United States), rabbit anti-p-Fascin-1 antibody (1:100, ab90618, Abcam, United States), mouse anti-Mas1 antibody (1:50, sc-390453, Santa Cruz, United States), rabbit antiCx3cr1 antibody (1:100, ab8021, Abcam, United States), mouse anti-Cx3cr1 antibody (1:50, sc-377227, Santa Cruz, United States), rat anti-GFAP antibody (1:100, 13–0300, Invitrogen, United States), rabbit anti-NeuN antibody (1:100, 24307, CST, United States), rabbit antiMBP antibody (1:100, AB5864, Merck, Germany), mouse anti-PKCα antibody (1:50, sc-8393, Santa Cruz, United States).

Techniques: Injection, Control, Solvent, Immunofluorescence