α scg10  (Novus Biologicals)

Journal: bioRxiv

Article Title: SARM1 is required for macrophage immunophenotype switching that is essential for nerve repair

doi: 10.64898/2026.04.07.716973

Figure Lengend Snippet: ( A ) Diagram of Mɸ-sensory neuron co-culture paradigm. Made in BioRender. ( B,C ) Representative images of WT DRGs cultured alone (DRG only) or with stimulated WT ( B ) or Sarm1-/- ( C ) Mɸ for 24 hours (mCSF, IL-4, or LPS). Scale bar = 250µm. ( D-F ) Quantification of DRG longest neurite length ( D ), total neurite length ( E ), or number of branch points per neuron of DRGs ( F ) in B , C . Error bars = SEM (N=≥90 neurons with WT Mɸ; ≥ 88 neurons with Sarm1-/- Mɸ ( E ) ≥93 neurons with WT Mɸ; ≥120 neurons with Sarm1-/- Mɸ ( F )≥63 neurons with WT Mɸs; ≥135 neurons with Sarm1-/- Mɸ from 3 independent experiments for WT Mɸ and 2 independent experiments for Sarm1-/- Mɸ. *p<0.05; **p<0.01; ****p<0.0001 by Kruskal-Wallis test with Dunn’s correction for multiple comparisons. ( G ) Representative images of WT or Sarm1-/- Mɸ injected into Sarm1-/- sciatic nerves. Images are 3 days after crush + injection. Mɸ are identified with F4/80 (green), and regenerating axons with SCG10 (Magenta). Arrows indicate regenerating axon tips. ( H,I ) Quantification of SCG10 expression following injection of Mɸ into Sarm1-/- ( H ) or WT ( I ) nerves. N=1-3 biological replicates. Expression was normalized to the injury site and represented as a fold change to the PBS control (black dashed line). Error bars = SEM.

Article Snippet: Primary antibodies diluted in blocking buffer were then added: chicken α-NFH (Aves Labs #NFH 1:100), chicken α-NFM (Aves Labs #NFM 1:100), chicken α-NFL (Aves Labs #NFL 1:100), rabbit α-CD68, (Cell Signaling Technology #E307V 1:500), rabbit α-CD206 (Cell Signaling Technology #E6T5J 1:500), α-SCG10 (Novus Biologicals #NBP1-49461 1:500), α-Myelin Basic Protein (Bio Legend #808401 1:500), α-ATP8A2 (Invitrogen #PA5 −65256 1:500) and/or rat α-F4/80 (Invitrogen #MA1-91124 1:500) and slides were incubated at 4°C overnight.

Techniques: Co-Culture Assay, Cell Culture, Injection, Expressing, Control

Journal: bioRxiv

Article Title: SARM1 is required for macrophage immunophenotype switching that is essential for nerve repair

doi: 10.64898/2026.04.07.716973

Figure Lengend Snippet: ( A,B ) Representative images of injury site ( A ) and distal stump ( B ) of sciatic nerves 7 days post SNC. Phosphotidyl serine flippase (Magenta; ATP8A2), Mɸ (Green; F4/80), and myelin (Yellow; MBP). Arrows indicate F4/80 and ATP8A2 positive cells. Scale bar = 50 µm. N = 3 biological replicates. ( C ) Quantification of images in ( A,B ). ATPA82 and MBP were normalized to an uninjured WT nerve. F4/80 signal was measured as intensity/area. N=2-3 biological replicates. Data is mean +/- SEM. *p<0.05 by two-way ANOVA with Dunnett correction for multiple comparisons. ( D ) Representative images of regenerating sensory axons (SCG10; Magenta) at 7d post SNC. N=3 biological replicates. Scale = 500 µm. ( E ) Quantification of relative SCG10 intensity from ( D ) normalized to injury site. Symbols indicate mean and shaded areas indicate SEM. N= 3 biological replicates. ( F-I ) Behavior analyses from BlackBox at 10 timepoints post unilateral SNC. Data in F-H are represented as mean +/- SEM. ( F ) Walking distance recorded in pixels. ( G ) Toe spread ratio of injured hindlimb to uninjured hindlimb. ( H ) Time spent rearing over 20 minutes recorded in seconds. ( I ) Weight bearing ratio of injured to uninjured hind paw. 1 indicates the most pressure recorded. *p<0.05; **p<0.01; ***p<0.005; ****p<0.0001 by two-way ANOVA with Dunnett correction for multiple comparisons. Biological replicates (WT = 10; mac-cKO = 13; neu-cKO =11; Sarm1-/- = 12)

Article Snippet: Primary antibodies diluted in blocking buffer were then added: chicken α-NFH (Aves Labs #NFH 1:100), chicken α-NFM (Aves Labs #NFM 1:100), chicken α-NFL (Aves Labs #NFL 1:100), rabbit α-CD68, (Cell Signaling Technology #E307V 1:500), rabbit α-CD206 (Cell Signaling Technology #E6T5J 1:500), α-SCG10 (Novus Biologicals #NBP1-49461 1:500), α-Myelin Basic Protein (Bio Legend #808401 1:500), α-ATP8A2 (Invitrogen #PA5 −65256 1:500) and/or rat α-F4/80 (Invitrogen #MA1-91124 1:500) and slides were incubated at 4°C overnight.

Techniques:

Journal: bioRxiv

Article Title: Sarm1 is not necessary for activation of neuron-intrinsic growth programs yet required for the Schwann cell repair response and peripheral nerve regeneration

doi: 10.1101/2024.03.04.583374

Figure Lengend Snippet: ( A ) Longitudinal sections of 3 dpc WT and Sarm1-/- sciatic nerves stained with anti-SGC10. Yellow dotted lines mark the injury site. Scale bar, 500 µm. ( B ) Longitudinal sections of 7 dpc WT and Sarm1-/- sciatic nerves stained with anti-SGC10. Yellow dotted lines mark the injury site. Scale bar, 500 µm. ( B’ ) Higher magnification images of SCG10 labeled 7dpc nerves, at the injury site, 3 mm, and 7 mm distal to the injury site. Scale bar, 100 µm. ( C ) Schematic of lumbar spinal cord, DRGs, and sciatic nerve. Brackets depict nerve segments microdissected for Western blotting. ( D ) Immunoblots of nerve segments showing SARM1 and SCG10 in the proximal (Prox), injury site (Inj), and distal (Dist) segments of WT and Sarm1-/- mice at 3 and 7 dpc (n = 5). Anti-ERK1/2 was used as a loading control. ( E and F ) Quantification of immunoblots shown in D; n = 5 mice per genotype and time point. Data normalized to ERK1/2 and shown as fold change from proximal nerve for each biological replicate (one-way ANOVA; **** p < 0.0001; ns = not significant).

Article Snippet: For staining the following primary antibodies were used: rabbit α-STMN2/ SCG10 (Novus Biologicals, NBP149461, 1:2000), α-STMN2/SCG10 (Thermo Fisher Scientific, 720178, 1:1000); rat α-F4/80 (Thermo Fisher Scientific ma1-91124, 1:500), CD68, α-Iba1 (Wako Chemicals, 019-19741, 1:1000), goat α-p75 NTR (Neuromics GT15057, 1:1000); chicken α-neuofilament H (Aves Lab NFH, 1:1000), and c-Jun (Cell Signaling #9165, 1:200), in blocking buffer overnight at 4°C in a humidified chamber.

Techniques: Staining, Labeling, Western Blot

Journal: bioRxiv

Article Title: Sarm1 is not necessary for activation of neuron-intrinsic growth programs yet required for the Schwann cell repair response and peripheral nerve regeneration

doi: 10.1101/2024.03.04.583374

Figure Lengend Snippet: ( A ) Timeline of the double (d)SNC lesion paradigm. Ten days after the first SNC, a second lesion was placed immediately proximal, and nerves harvested 3d after the second lesion. ( B ) Longitudinal sections of dSNC WT and Sarm1-/- sciatic nerves showing the injury site and nerve segments 3 mm and 5 mm distal from the injury site, stained for p75 NTR (red) and c-Jun (green). Scale bar, 100 µm. ( C ) Western blots of nerve segments probed for p75 NTR , c-Jun, and SARM1. WT and Sarm1-/- nerves from sham-operated mice, dSNC injury site and distal nerve are shown. ERK1/2 was used as loading control. ( D, E ) Quantification of Western blots shown in (H); 5 biological replicates with different color shades. Data was normalized to ERK1/2 and shown as fold change from sham-operated WT or Sarm1-/- nerves (one-way ANOVA; * p < 0.05; ** p < 0.01), ns = not significant. ( F ) Anti-NF-H labeled DRG neurons from naïve WT mice co-cultured with SC harvested from WT and Sarm1-/- dSNC distal nerves. Scale bar, 250 µm. ( G ) Quantification of DRG neurite length in cultures shown in (D); WT, 188 cells (n= 3); Sarm1-/- , 146 cells (n= 3). Neurite length ± SEM is shown. Student’s t-test, ns = not significant. ( H ) Longitudinal sections of 3d SNC (single crush) WT and Sarm1-/- nerves stained with anti-SCG10. The yellow dotted lines denote the injury site. Scale bar, 500 µm. ( H’ ) Longitudinal sections of dSNC nerves of WT and Sarm1-/- mice stained with anti-SCG10. The second crush site is marked with white dotted lines. Scale bar, 500 µm ( I ) Higher magnification images of SCG10 labeled dSNC WT and Sarm1-/- mice at 3 and 5 mm distal to the injury site. Scale bar, 100 µm. ( J ) Quantification of SCG10 mean fluorescence intensity (MFI) measured at 500 µm intervals from the injury site. Yellow box denotes the first crush site in the dSNC paradigm or the single crush site for 3d SNC; a , WT 3 dpc vs Sarm1-/- 3 dpc (single crush); b , WT 3 dpc vs WT dSNC; c, WT dSNC vs Sarm1-/- dSNC. MFI ± SEM is shown. Number of a , b , or c letters is equivalent to the number of asterisks (* p < 0.05; ** p < 0.01; *** p < 0.001). N = 3 mice per group. ( K ) Longitudinal assessment of toe spreading reflex; mean distance between the tips of the first and fifth toes ± SEM is shown. N = 7 mice per genotype and time point. Two-way ANOVA; ** p < 0.01; *** p < 0.001; **** p < 0.0001; ns = not significant.

Article Snippet: For staining the following primary antibodies were used: rabbit α-STMN2/ SCG10 (Novus Biologicals, NBP149461, 1:2000), α-STMN2/SCG10 (Thermo Fisher Scientific, 720178, 1:1000); rat α-F4/80 (Thermo Fisher Scientific ma1-91124, 1:500), CD68, α-Iba1 (Wako Chemicals, 019-19741, 1:1000), goat α-p75 NTR (Neuromics GT15057, 1:1000); chicken α-neuofilament H (Aves Lab NFH, 1:1000), and c-Jun (Cell Signaling #9165, 1:200), in blocking buffer overnight at 4°C in a humidified chamber.

Techniques: Staining, Western Blot, Labeling, Cell Culture, Fluorescence

Journal: bioRxiv

Article Title: Sarm1 is not necessary for activation of neuron-intrinsic growth programs yet required for the Schwann cell repair response and peripheral nerve regeneration

doi: 10.1101/2024.03.04.583374

Figure Lengend Snippet: ( A ) Schematic of the sciatic nerve grafting experiment. A WT or Sarm1-/- nerve segment was grafted into a WT host. Coaptation of the graft was done only at the proximal end. Intraoperative and harvested graft images are shown. Scale bar, 1 mm. ( B ) Representative longitudinal nerve sections showing WT and a Sarm1-/- grafts (dotted lines) after 14 days, stained with fluoromyelin and anti-p75 NTR . High magnification images on the right show difference in myelin integrity between WT and Sarm1-/- grafts. Scale bar for low magnification, 500 µm; scale bar for high magnification, 100 µm. ( C ) Nerve sections stained with anti-SCG10 and anti-p75 NTR of WT and Sarm1-/- grafts. ( D ) High magnification of WT and Sarm1-/- grafts stained with p75 NTR and SCG10. Scale bar, 100 µm. ( E and F ) Quantification of p75 NTR and SCG10 mean fluorescence intensity ± SEM at 500 µm intervals within the grafts (n= 6 biological replicates per genotype).

Article Snippet: For staining the following primary antibodies were used: rabbit α-STMN2/ SCG10 (Novus Biologicals, NBP149461, 1:2000), α-STMN2/SCG10 (Thermo Fisher Scientific, 720178, 1:1000); rat α-F4/80 (Thermo Fisher Scientific ma1-91124, 1:500), CD68, α-Iba1 (Wako Chemicals, 019-19741, 1:1000), goat α-p75 NTR (Neuromics GT15057, 1:1000); chicken α-neuofilament H (Aves Lab NFH, 1:1000), and c-Jun (Cell Signaling #9165, 1:200), in blocking buffer overnight at 4°C in a humidified chamber.

Techniques: Staining, Fluorescence