anti scg10  (Novus Biologicals)

Journal: bioRxiv

Article Title: Herpes simplex virus pUL56 abolishes neuronal activity by removing voltage-gated ion channels from the plasma membrane

doi: 10.64898/2026.04.10.717620

Figure Lengend Snippet: ( A ) Single-step (MOI 5) replication curve of wildtype (WT) and ΔpUL56 HSV-1 in day 14 i3Neurones. Mean ± SD for three independent experiments is shown. ( B ) Schematic of the QTV workflow using day 14 i3Neurones. Mock samples were harvested at 12 hpi for all analyses, and additionally at 3 and 30 hpi for transcriptomics (RNAseq) using Oxford Nanopore Technology (ONT). ( C ) Changes in i3Neurone protein abundance at 30 hpi with WT HSV-1. Horizontal axis shows average fold change and vertical axis shows significance (two-sided t-test) for three independent experiments. Proteins are coloured by FDR-corrected significance and viral proteins are outlined in pink. ( D ) Changes in i3Neurone transcript abundance at 30 hpi with WT HSV-1. Horizontal axis shows average fold change and vertical axis shows FDR-adjusted significance for three independent experiments. Significantly altered genes (log 2 fold change ≥ 2 and p ≤ 0.05) are blue and viral transcripts are outlined in pink. ( E ) Immunoblot analysis of day 14 i3Neurones at 16 hpi (MOI 10) with HSV-1 strains KOS, S17 and SC16, or Semliki Forest virus (SFV)4. Changes in KIF1A, cFOS and GOPC are HSV-1 specific, whereas abundance of stathmins (STMN2–4) is altered in all infections. Capsid proteins VP5 (HSV) and CP (SFV4) confirm successful infection and GAPDH is a loading control. ( F ) Gene Ontology (GO) analysis of proteins (top, blue) and transcripts (bottom, green) with significantly changed abundance at 30 hpi in HSV-versus mock-infected i3Neurones. Significant GO molecular function terms are shown as bubbles sized by magnitude of enrichment, with significance (FDR-adjusted p-value) plotted on the horizontal axis and arrows indicating whether the term was enriched amongst the up- or down-regulated gene products. ( G ) Changes in protein (horizontal) versus transcript (vertical) abundance in mock-versus HSV-infected i3Neurones at 30 hpi. Cellular and viral gene products are grey and pink, respectively, with selected cellular gene products highlighted (black). Potential targets of HSV-1 directed proteasomal degradation (larger fold change in protein vs transcript abundance) are to the left of the dotted red line. (H) Normalised abundance traces of selected proteins (top) and transcripts (bottom) across the time course of infection with WT (pink) or ΔpUL56 (blue) HSV-1. Mock-infected samples (M) are shown as 0 hpi. ( I ) Cellular protein abundance changes in day 14 i3Neurones infected (MOI 5) with WT versus ΔpUL56 i3Neurones. Data are plotted as in ( C ), with horizontal axis showing average mock-corrected fold change for three independent experiments.

Article Snippet: Primary and secondary antibody incubations were performed in TBS supplemented with 0.1% TWEEN-20 and 5% (w/v) skim milk powder using the following antibodies: HSV-1 VP5, ( ); SV4 CP, kindly provided by Andres Merits ( ); KIF1A, BD Transduction Laboratories 612094; cFOS, Cell Signalling Technologies 2250T; GOPC, Abcam Ab133472; STMN2, Novus Biologicals NBP1-49461; STMN3, ProteinTech 11311-1-AP; STMN4, ProteinTech 12027-1-AP; GAPDH, GeneTex GTX28245; HSV-1 pUL56, ( ); HA, Cell Signalling Technologies C29F4; Anti-rabbit 800, LI-COR 926-32213; Anti-rabbit 680, LI-COR 926-68023; Anti-mouse 800, LI-COR 926-32210; Anti-mouse 680, LI-COR 926-68020.

Techniques: Transcriptomics, RNA sequencing, Quantitative Proteomics, Western Blot, Virus, Infection, 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 ) 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: