anti scg 10  (Novus Biologicals)

Journal: Frontiers in Cellular Neuroscience

Article Title: A Conditioning Sciatic Nerve Lesion Triggers a Pro-regenerative State in Primary Sensory Neurons Also of Dorsal Root Ganglia Non-associated With the Damaged Nerve

doi: 10.3389/fncel.2019.00011

Figure Lengend Snippet: Representative pictures of cryostat sections through the DRG from naïve (A,G) , sham-operated (B,H) rats and rats with unilateral SNC (C,D,I,J) or CSNT (E,F,K,L ) for 7 days. The sections of ipsilateral (C,E) and contralateral (D,F) DRG of the L4 spinal segment as well as ipsilateral (I,K) and contralateral (J,L) DRG of the C7 spinal segment were incubated under the same conditions with rabbit polyclonal antibody recognizing superior cervical ganglion-10 (SCG-10). Scale bars = 75 μm. (M) Graph illustrating the mean intensity of SCG-10 immunofluorescence measured in individual DRG neuron-size classes of cervical (C6-C8) and lumbar (L4-L5) spinal segments ipsilateral (i) and contralateral (c) to unilateral SNC and transection (CSNT) for 7 days; n = 6 for each group. Neurons A ≥ 40 μm; Neurons B 25–40 μm; Neurons C ≤ 25 μm. *Significant difference ( p < 0.05) compared to naive or sham-operated rats in a Mann-Whitney U-test.

Article Snippet: Blots were blocked using 1% BSA in PBST (3.2 mM Na 2 HPO 4 , 0.5 mM KH 2 PO 4 , 1.3 mM KCl, 135 mM NaCl, 0.05% Tween 20, pH 7.4) for 1 h and incubated with anti-GAP-43 mouse monoclonal (1:1,000; Sigma, Ronkonkoma, NY, USA), rabbit polyclonal anti-phosphorylated S41-GAP43 (1:500; Thermo Fisher Scientific, Waltham, MA, USA) or anti-SCG-10 (1:500; LSBio, Seattle, WA, USA) antibodies overnight.

Techniques: Incubation, Immunofluorescence, MANN-WHITNEY

Journal: Frontiers in Cellular Neuroscience

Article Title: A Conditioning Sciatic Nerve Lesion Triggers a Pro-regenerative State in Primary Sensory Neurons Also of Dorsal Root Ganglia Non-associated With the Damaged Nerve

doi: 10.3389/fncel.2019.00011

Figure Lengend Snippet: Results of western blot analysis of GAP-43, pGAP-43 and SCG-10 protein levels in DRG of L4-L5 (L) and C6-C8 (C) segments removed from ipsilateral (i) and contralateral (c) sides of naïve as well as sham-, SNC- and CSNT-operated rats for 7 days. Upper panel (A) illustrates representative western blots of DRG from three rats for each group. Equal loading of proteins was confirmed by actin levels (Actin). The same Actin controls were used for analysis of GAP-43, pGAP-43 and SCG-10 protein levels in this set representative western blots. Lower panels (B) show densitometry of individual protein bands after normalization to actin from three independent experiments; the intensities of the bands from naïve DRG were as arbitrarily set to 1. *Significant difference ( p < 0.05) when compared to sham-operated rats; † Significant difference ( p < 0.05) compared to contralateral DRG in a Mann-Whitney U-test.

Article Snippet: Blots were blocked using 1% BSA in PBST (3.2 mM Na 2 HPO 4 , 0.5 mM KH 2 PO 4 , 1.3 mM KCl, 135 mM NaCl, 0.05% Tween 20, pH 7.4) for 1 h and incubated with anti-GAP-43 mouse monoclonal (1:1,000; Sigma, Ronkonkoma, NY, USA), rabbit polyclonal anti-phosphorylated S41-GAP43 (1:500; Thermo Fisher Scientific, Waltham, MA, USA) or anti-SCG-10 (1:500; LSBio, Seattle, WA, USA) antibodies overnight.

Techniques: Western Blot, MANN-WHITNEY

Journal: Frontiers in Cellular Neuroscience

Article Title: A Conditioning Sciatic Nerve Lesion Triggers a Pro-regenerative State in Primary Sensory Neurons Also of Dorsal Root Ganglia Non-associated With the Damaged Nerve

doi: 10.3389/fncel.2019.00011

Figure Lengend Snippet: Results of real-time PCR (RT-PCR) of relative GAP-43 (A) and SCG-10 (B) mRNA levels in DRG of lumbar (L4-L5) and cervical (C6-C8) spinal segments of removed from ipsilateral (i) and contralateral (c) sides of Naïve as well as sham-, SNC- and CSNT-operated rats for 7 days; n = 6 for each group. Relative expressions were determined using Actin as the housekeeping gene and normalized to naïve controls. *Significant difference ( p < 0.05) compared to sham-operated rats in a Mann-Whitney U-test.

Article Snippet: Blots were blocked using 1% BSA in PBST (3.2 mM Na 2 HPO 4 , 0.5 mM KH 2 PO 4 , 1.3 mM KCl, 135 mM NaCl, 0.05% Tween 20, pH 7.4) for 1 h and incubated with anti-GAP-43 mouse monoclonal (1:1,000; Sigma, Ronkonkoma, NY, USA), rabbit polyclonal anti-phosphorylated S41-GAP43 (1:500; Thermo Fisher Scientific, Waltham, MA, USA) or anti-SCG-10 (1:500; LSBio, Seattle, WA, USA) antibodies overnight.

Techniques: Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, MANN-WHITNEY

Journal: Frontiers in Cellular Neuroscience

Article Title: A Conditioning Sciatic Nerve Lesion Triggers a Pro-regenerative State in Primary Sensory Neurons Also of Dorsal Root Ganglia Non-associated With the Damaged Nerve

doi: 10.3389/fncel.2019.00011

Figure Lengend Snippet: (A) Representative longitudinal sections through the ulnar nerves distal to the crush site (solid line) showing SCG-10 positive regenerated axons. Arrows indicate the tip of the longest SCG-10 positive axons. The ulnar nerves were removed after 1 day from control rat (only ulnar nerve crush) and from rats 7 days after prior SNC or CSNT. Scale bars = 100 μm. (B) The top portion of the graph illustrates mean length of regenerated SCG10+ axons ± SD in the ulnar nerve 1 day after crush in rats without a sciatic nerve injury (Control) and following 7 days from SNC or CSNT; n = 6 for each group. *Significant difference ( p < 0.05) compared to control. The middle portion illustrates mean length of regenerated SCG10+ axons ± SD in the ulnar nerve 1 day after crush and intrathecal injection of 10 μl of artificial cerebrospinal fluid (ACSF) or IL-6 (20 ng/10 μl); n = 4 for each group. **Significant difference ( p < 0.05) compared to control. The bottom portion illustrates mean length of regenerated SCG10+ axons ± SD in the ulnar nerve 1 day after crush 7 days from prior CSNT and intrathecal application of ACSF (CSNT+ACSF) or JAK2 inhibitor (CSNT+AG490); n = 4 for each group. ‡ Significant difference ( p < 0.05) compared to CSNT+ACSF group in a Mann-Whitney U-test.

Article Snippet: Blots were blocked using 1% BSA in PBST (3.2 mM Na 2 HPO 4 , 0.5 mM KH 2 PO 4 , 1.3 mM KCl, 135 mM NaCl, 0.05% Tween 20, pH 7.4) for 1 h and incubated with anti-GAP-43 mouse monoclonal (1:1,000; Sigma, Ronkonkoma, NY, USA), rabbit polyclonal anti-phosphorylated S41-GAP43 (1:500; Thermo Fisher Scientific, Waltham, MA, USA) or anti-SCG-10 (1:500; LSBio, Seattle, WA, USA) antibodies overnight.

Techniques: Injection, MANN-WHITNEY