rat anti-bmp10 antibody  (Novus Biologicals)

Journal: Frontiers in Pharmacology

Article Title: BMP10 accelerated spinal astrocytic activation in neuropathic pain via ALK2/smad1/5/8 signaling

doi: 10.3389/fphar.2024.1426121

Figure Lengend Snippet: Graphical representation of the experimental design. Experiment 1: Changes in pain sensitization and expressions of BMP10, ALK2, Smad1/5/8, phosphorylated Smad1/5/8, and GFAP after SNI in mice. Experiment 2: The effects of BMP10 siRNA on SNI-induced pain hypersensitivity and astrocytic activation. Experiment 3: The involvements of ALK2 in BMP10-induced pain hypersensitivity and astrocytic activation. Experiment 4: The effects of Smad1 siRNA on BMP10-induced pain hypersensitivity and astrocytic activation.

Article Snippet: Subsequently, the tissues were incubated with the primary antibodies for 12 h at 4°C as follows: mouse anti-GFAP antibody (1:200, ab279289, Abcam), mouse anti-CGRP antibody (1:500, ab81887, Abcam), mouse anti-c-fos antibody (1:1,000, ab208942, Abcam), mouse anti-Iba-1 antibody (1:200, ab283319, Abcam), rat anti-BMP10 antibody (1:200, 654319, Novus Biologicals, CO, United States) and rabbit anti-ALK2 antibody (1:100, PA5-114818, Thermo Fisher Scientific, MA, United States).

Techniques: Activation Assay

Journal: Frontiers in Pharmacology

Article Title: BMP10 accelerated spinal astrocytic activation in neuropathic pain via ALK2/smad1/5/8 signaling

doi: 10.3389/fphar.2024.1426121

Figure Lengend Snippet: Expression levels of BMP10, GFAP in mouse ipsilateral spinal dorsal horn after SNI. (A, B) Western blot analysis showed the expression levels of BMP10 and GFAP in the ipsilateral (above) and contralateral (below) spinal dorsal horn of sham and SNI mice; (C, D) Immunofluorescence staining showed the expression of GFAP in the ipsilateral and contralateral spinal dorsal horn of sham and SNI mice (scale bar = 200 μm/50 μm); (E–G) Double immunofluorescence staining showed the coexpression of BMP10 (red) with GFAP (green) in the ipsilateral spinal dorsal horn of sham and SNI mice on postoperative Day 14 (scale bar = 200 μm/50 μm). Data are presented as mean and SEM; sham mice versus SNI mice on postoperative Days 7 and 14, *** p < 0.001 (n = 6).

Article Snippet: Subsequently, the tissues were incubated with the primary antibodies for 12 h at 4°C as follows: mouse anti-GFAP antibody (1:200, ab279289, Abcam), mouse anti-CGRP antibody (1:500, ab81887, Abcam), mouse anti-c-fos antibody (1:1,000, ab208942, Abcam), mouse anti-Iba-1 antibody (1:200, ab283319, Abcam), rat anti-BMP10 antibody (1:200, 654319, Novus Biologicals, CO, United States) and rabbit anti-ALK2 antibody (1:100, PA5-114818, Thermo Fisher Scientific, MA, United States).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining, Double Immunofluorescence Staining

Journal: Frontiers in Pharmacology

Article Title: BMP10 accelerated spinal astrocytic activation in neuropathic pain via ALK2/smad1/5/8 signaling

doi: 10.3389/fphar.2024.1426121

Figure Lengend Snippet: Intrathecal injection of BMP10 siRNA decreased the expression of BMP10 without conspicuous side effects. (A) qRT-PCR analysis showed the expression levels of BMP10 mRNA in the spinal cord of mice after intrathecal injection with the vehicle, NC siRNA, and BMP10 siRNA; (B, C) Western blot analysis showed the expression levels of BMP10 protein in the spinal cord of mice after intrathecal injected with vehicle, NC siRNA, BMP10 siRNA; (D, E) Paw withdrawal threshold (PWT) and thermal withdrawal latency (TWL) of mice after intrathecal injected with vehicle, NC siRNA, BMP10 siRNA; (F) Representative movement traces in OFT tests of mice after intrathecal injected with vehicle, NC siRNA, BMP10 siRNA (G–I) Time in center zone, average speed, and total distance of mice after intrathecal injected with vehicle, NC siRNA, BMP10 siRNA. Data were presented as mean and SEM, BMP10 siRNA group versus Control group ** p < 0.01, *** p < 0.001; BMP10 siRNA group versus NC siRNA group ## p < 0.01, ### p < 0.001, n = 6.

Article Snippet: Subsequently, the tissues were incubated with the primary antibodies for 12 h at 4°C as follows: mouse anti-GFAP antibody (1:200, ab279289, Abcam), mouse anti-CGRP antibody (1:500, ab81887, Abcam), mouse anti-c-fos antibody (1:1,000, ab208942, Abcam), mouse anti-Iba-1 antibody (1:200, ab283319, Abcam), rat anti-BMP10 antibody (1:200, 654319, Novus Biologicals, CO, United States) and rabbit anti-ALK2 antibody (1:100, PA5-114818, Thermo Fisher Scientific, MA, United States).

Techniques: Injection, Expressing, Quantitative RT-PCR, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: BMP10 accelerated spinal astrocytic activation in neuropathic pain via ALK2/smad1/5/8 signaling

doi: 10.3389/fphar.2024.1426121

Figure Lengend Snippet: Intrathecal injection of BMP10 siRNA inhibited the SNI-induced pain hypersensitivity and astrocytic activation. (A, B) The time course of PWT and TWL in sham and SNI mice after intrathecal delivery of siRNAs; Data are presented as mean and SEM; SNI + NC siRNA group and SNI + BMP10 siRNA group versus Sham + NC siRNA *** p < 0.001; SNI + BMP10 siRNA versus SNI + NC siRNA group # p < 0.05 ## p < 0.01 (n = 12); (C, D) Western blot showed the expression levels of BMP10 and GFAP in the spinal cord of sham and SNI mice after intrathecal delivery of siRNAs on postoperative Day 14; (E, F) Immunofluorescence staining results showing the expression of GFAP in the spinal dorsal horn of sham and SNI mice after intrathecal delivery of siRNAs on postoperative Day 14 (scale bar = 100 μm/50 μm); Data are presented as mean and SEM; SNI + NC siRNA group and SNI + BMP10 siRNA group versus Sham + NC siRNA*** p < 0.001; SNI + BMP10 siRNA versus SNI + NC siRNA group # p < 0.05, ### p < 0.001 (n = 6).

Article Snippet: Subsequently, the tissues were incubated with the primary antibodies for 12 h at 4°C as follows: mouse anti-GFAP antibody (1:200, ab279289, Abcam), mouse anti-CGRP antibody (1:500, ab81887, Abcam), mouse anti-c-fos antibody (1:1,000, ab208942, Abcam), mouse anti-Iba-1 antibody (1:200, ab283319, Abcam), rat anti-BMP10 antibody (1:200, 654319, Novus Biologicals, CO, United States) and rabbit anti-ALK2 antibody (1:100, PA5-114818, Thermo Fisher Scientific, MA, United States).

Techniques: Injection, Activation Assay, Western Blot, Expressing, Immunofluorescence, Staining

Journal: Frontiers in Pharmacology

Article Title: BMP10 accelerated spinal astrocytic activation in neuropathic pain via ALK2/smad1/5/8 signaling

doi: 10.3389/fphar.2024.1426121

Figure Lengend Snippet: Intrathecal injection of BMP peptide evoked pain hypersensitivity and astrocytic activation in normal mice (A, B) The time course of PWT and TWL in normal mice after intrathecal delivery of BMP10 peptide and vehicle; Data were presented as mean and SEM, 100 ng group and 1000 ng group versus Vehicle group * p < 0.05, ** p < 0.01, *** p < 0.001; n = 6; (C) Western blot showed the expression levels of GFAP in spinal cord of mice after intrathecal delivery of BMP10 peptide and vehicle; (D) Western blot showed the expression levels of BMP10 in spinal cord at different time points after intrathecal delivery of BMP10 peptide (100 ng); (E–H) Immunofluorescence staining results showing the expression of c-fos, CGRP, and Iba-1 in the spinal dorsal horn of mice after intrathecal delivery of BMP10 peptide and vehicle (scale bar = 200 μm/50 μm). Data were presented as mean and SEM, 100 ng group and 1,000 ng group versus Vehicle group ** p < 0.01, *** p < 0.001; n = 6.

Article Snippet: Subsequently, the tissues were incubated with the primary antibodies for 12 h at 4°C as follows: mouse anti-GFAP antibody (1:200, ab279289, Abcam), mouse anti-CGRP antibody (1:500, ab81887, Abcam), mouse anti-c-fos antibody (1:1,000, ab208942, Abcam), mouse anti-Iba-1 antibody (1:200, ab283319, Abcam), rat anti-BMP10 antibody (1:200, 654319, Novus Biologicals, CO, United States) and rabbit anti-ALK2 antibody (1:100, PA5-114818, Thermo Fisher Scientific, MA, United States).

Techniques: Injection, Activation Assay, Western Blot, Expressing, Immunofluorescence, Staining

Journal: Frontiers in Pharmacology

Article Title: BMP10 accelerated spinal astrocytic activation in neuropathic pain via ALK2/smad1/5/8 signaling

doi: 10.3389/fphar.2024.1426121

Figure Lengend Snippet: ALK2 was involved in BMP10-induced pain hypersensitivity and astrocytic activation. (A, B) The time course of PWT and TWL in normal mice after intrathecal delivery of BMP10 peptide and reagents; Data are presented as mean and SEM, Vehicle group versus BMP10 group * p < 0.05, ** p < 0.01, *** p < 0.001; BMP10+ALK2-IN-2 group versus BMP group # p < 0.05, ## p < 0.01, ### p < 0.001 (n = 12); (C) Representative movement traces in OFT tests of mice after intrathecal delivery of BMP10 peptide and reagents; (D–F) Time in center zone, average speed and total distance of mice in open field after intrathecal delivery of BMP10 peptide and reagents; (G, H) immunofluorescence staining showed the expression of GFAP in the spinal dorsal horn after intrathecal delivery of BMP10 peptide and reagents (scale bar = 200/50 μm); (I) Western blot showed the expression of GFAP in the spinal cord after intrathecal delivery of BMP10 peptide and reagents; Data are presented as mean and SEM, Vehicle group versus BMP10 group * p < 0.05, ** p < 0.01, *** p < 0.001; BMP10+ ALK2-IN-2 group versus BMP group # p < 0.05, ## p < 0.01, ### p < 0.001 (n = 6).

Article Snippet: Subsequently, the tissues were incubated with the primary antibodies for 12 h at 4°C as follows: mouse anti-GFAP antibody (1:200, ab279289, Abcam), mouse anti-CGRP antibody (1:500, ab81887, Abcam), mouse anti-c-fos antibody (1:1,000, ab208942, Abcam), mouse anti-Iba-1 antibody (1:200, ab283319, Abcam), rat anti-BMP10 antibody (1:200, 654319, Novus Biologicals, CO, United States) and rabbit anti-ALK2 antibody (1:100, PA5-114818, Thermo Fisher Scientific, MA, United States).

Techniques: Activation Assay, Immunofluorescence, Staining, Expressing, Western Blot

Journal: Frontiers in Pharmacology

Article Title: BMP10 accelerated spinal astrocytic activation in neuropathic pain via ALK2/smad1/5/8 signaling

doi: 10.3389/fphar.2024.1426121

Figure Lengend Snippet: Smad1 was involved in BMP10-induced pain hypersensitivity and astrocytic activation. (A, B) PWT and TWL of normal mice after intrathecal delivery of siRNA and BMP10 peptide; (C, D) Western blot showed the expressions of p-Smad1/5/8 and GFAP in the spinal cord of normal mice after intrathecal delivery of siRNA and BMP10 peptide; (E, F) immunofluorescence staining showed the expression of GFAP in the spinal dorsal horn of normal mice after the intrathecal delivery of siRNA and BMP10 peptide; (scale bar = 200 μm/50 μm); Data are presented as mean and SEM, NC siRNA + Vehicle group versus NC siRNA + BMP10 group *** p < 0.001; NC siRNA + BMP10 group versus Smad1 siRNA + BMP10 group # p < 0.05, ### p < 0.001 (n = 6).

Article Snippet: Subsequently, the tissues were incubated with the primary antibodies for 12 h at 4°C as follows: mouse anti-GFAP antibody (1:200, ab279289, Abcam), mouse anti-CGRP antibody (1:500, ab81887, Abcam), mouse anti-c-fos antibody (1:1,000, ab208942, Abcam), mouse anti-Iba-1 antibody (1:200, ab283319, Abcam), rat anti-BMP10 antibody (1:200, 654319, Novus Biologicals, CO, United States) and rabbit anti-ALK2 antibody (1:100, PA5-114818, Thermo Fisher Scientific, MA, United States).

Techniques: Activation Assay, Western Blot, Immunofluorescence, Staining, Expressing