Journal: Journal of Neuroscience

Article Title: Toll-Like Receptor Signaling Is Critical for Wallerian Degeneration and Functional Recovery after Peripheral Nerve Injury

doi: 10.1523/jneurosci.3027-07.2007

Figure Lengend Snippet: Figure 3. Deficiency in TLR signaling compromises macrophage recruitment/activation and delays myelin debris clearance, axonal regeneration, and locomotor recovery after sciatic nerve lesion. A–D, Representative fluorescence photomicrographs takenfromlongitudinalsectionsofthesciaticnervedistaltothelesionshowingCD68-immunopositivemacrophagesinC57BL/6J (A),TLR2-ko(B),C3H/HeOUJ(C),andTLR4d(D)miceat7d.E,QuantificationofthenumberofCD68macrophagesinthesciaticnerve distalstumpofTLR2-ko,TLR4d,MyD88-ko,andwild-typemiceat7dafterlesion(n8pergroup).F–I,Bright-fieldphotomicrographs showingmyelinstainedwithLFBinthesciaticnervedistalstumpofC57BL/6J(F),TLR2-ko(G),C3H/HeOUJ(H),andTLR4d(I)miceat7d. J,QuantificationofLFBstainingofmyelininthedegeneratingsciaticnervedistalstumpofmicedeficientinTLRsignalingat7d(n8per group). K, Quantification of the number of axons that had regenerated up to 4 mm distal to the site of lesion, as visualized by GAP-43 immunofluorescence, at 4 d after lesion. L, M, Recovery of locomotor functions from a sciatic nerve lesion, as determined by the SFI, in TLR2-ko,MyD88-ko,andTLR4dmicecomparedwiththeirrespectivecontrolgroups(WT)overaperiodof49d(n12pergroup).***p 0.001,**p0.01,and*p0.05comparedwiththecontrolgroup.Scalebar,50m.

Article Snippet: Immunohistochemistry was performed to detect the following antigens: (1) mouse CD68 using the monoclonal anti-CD68 antibody (to visualize activated macrophages/monocytes; Serotec, Raleigh, NC), (2) growth-associated protein-43 (GAP-43) using the polyclonal antiGAP-43 antibody (to visualize regenerating axons; Novus Biologicals, Littleton, CO), (3) rat CD68 using the monoclonal ED-1 antibody (to visualize activated macrophages/monocytes; Serotec), (4) ionized calcium-binding adaptor molecule 1 (iba1) using the polyclonal antiiba1 antibody (to visualize macrophages/monocytes; Wako Chemicals USA, Richmond, VA), and (5) mouse CD45 using the monoclonal antiCD45 antibody (to visualize leukocytes; BD PharMingen, Mississauga, Ontario, Canada).

Techniques: Activation Assay, Fluorescence, Immunofluorescence

Journal: bioRxiv

Article Title: Adipose mTORC2 is essential for arborization of sensory neurons in white adipose tissue and whole-body energy homeostasis

doi: 10.1101/2022.03.21.485116

Figure Lengend Snippet: (A) Immunoblot analysis of inguinal WAT (iWAT) tissue from control and iAdRiKO mice two weeks after tamoxifen treatment. (n=6;6). (B) Immunoblot analysis of iWAT tissue from control and iAdRiKO mice four weeks after tamoxifen treatment. (n=6;6). (C) Immunoblot analysis of surgically denervated iWAT depot (denervation) compared to iWAT depot from sham-operated mice (sham). Neurofilament heavy polypeptide (NFH). (n=5;5). (D) Representative image of a large nerve bundle in iWAT of control mice immunostained with growth-associated protein 43 (GAP43)-pS41 and calcitonin gene-related peptide (CGRP). (N=11;9). (E) Representative image of a large nerve bundle in iWAT of control mice immunostained with GAP43-pS41 and tyrosine hydroxylase (TH). (N=19;11).

Article Snippet: Primary antibodies used were RICTOR (1:1000; Cell signaling; Cat#2140), AKT (1:1000; Cell signaling, Cat#4685), AKT-pS473 (1:1000; Cell signaling, Cat#9271), CALNEXIN (1:1000, Enzo, Cat#ADI-SPA-860-F), GAP43 (1:1000, Cell signaling, Cat#8945), GAP43-pS41 (1:1000, R&D Systems, Cat#PPS006), Tyrosine hydroxylase (1:500, Millipore, Cat#AB1542), HSL (1:2000, Cell signaling, Cat#4107), HSL-pS660 (1:1000, Cell signaling, Cat#4126), HSL-pS563 (1:1000, Cell signaling, Cat#4139).

Techniques: Western Blot, Control