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: