Journal: Cells
Article Title: Chronic Pain Induced by Social Defeat Stress in Juvenile Mice Depends on TLR4
doi: 10.3390/cells14050350
Figure Lengend Snippet: SDS increases TLR4 dimers of spinal cord microglia and DRG macrophages. ( A ) TLR4 dimers (%) in SDS and no-SDS groups of microglia (CD11b + population) and ( B ) macrophages (CD11b + /F4-80 + population) on day 22 of the protocol. ( C ) Gating strategy for lumbar spinal cord cells, first live cells were gated (APC-Cy7 live/dead Ghost dye); second, microglia population (PercpCy 5.5-conjugated CD11b+), lastly—TLR4 monomers (PC7-conjugated TLR4/MD2) or TLR4 total (APC-conjugated total TLR4). ( D ) Gating strategy for dorsal root ganglia cells, first live cells were gated (APC-Cy7 live/dead Ghost dye); second, microglia population (PercpCy 5.5-conjugated CD11b+), third macrophages population (488-conjugated F4-80) and lastly—TLR4 monomers (PC7-conjugated TLR4/MD2) or TLR4 total (APC-conjugated total TLR4). Data represent mean ± SEM, t -test, SDS vs. no-SDS, * p < 0.05, ** p < 0.01, n = 6 (SDS), and n = 6 (no-SDS).
Article Snippet: Single-cell suspensions were incubated for 45 min on ice with the following antibodies: APC-Cy7 live/dead Ghost dye (#18452, Cell Signaling, Danvers, MA, USA) or 540 live/dead Ghost dye (#72086, Cell Signaling, Danvers, MA, USA); PC7-conjugated TLR4/MD2 (#MTS510; ThermoFisher, Waltham, MA, USA); APC-conjugated total TLR4 (#SA15-21; Biolegend, San Diego, CA, USA); 488-conjugated F4-80 (#123110; Biolegend, San Diego, CA, USA); and PercpCy 5.5-conjugated CD11b (#101228; Biolegend, San Diego, CA, USA).
Techniques:
