Journal: Nature Communications

Article Title: Targeting prostaglandin E 2 receptor 2 in Schwann cells inhibits inflammatory pain but not inflammation

doi: 10.1038/s41467-025-63782-8

Figure Lengend Snippet: a , d , f Dose-dependent non-evoked nociception and b , e , g dose- and time-dependent allodynia after intraplantar (i.pl.) injection of PGE 2 , L-902,688 (L-902), butaprost (Buta) or vehicle (Veh) in C57BL/6 J mice (B6) ( n = 8 mice/group). c Schematic representation of agonists/antagonists targeting EP2 and EP4 receptors. h Non-evoked nociception and i allodynia after i.pl. PGE 2 (1.5 nmol) or Veh in B6 pretreated with PF-04448948 (PF, 5nmol), BGC 20-1531 (BGC, 5 nmol) or Veh ( n = 8 mice/group). j RT-qPCR for Ptger2, Ptger4 Avil and S100 mRNA in mouse dorsal root ganglia (DRG) sciatic and cutaneous Schwann cells (SCs) (DRG and sciatic SCs n = 4, cutaneous n = 3 independent experiments). k , l Representative images of EP2, EP4, NeuN and S100B expression in mouse DRG and sciatic nerve tissue (scale bar: 20 μm) ( n = 3 subjects). m Representative images of EP2, EP4 and SOX10 expression in mouse sciatic and cutaneous SCs ( n = 3 independent experiments). n–r Non-evoked nociception (left panel) and allodynia (right panel) after i.pl. PGE 2 or Veh in Plp-Cre, Adv-Cre or Control mice infected with AAV for selective silencing of EP4 (-Ptger4 ) ( P1p-Ptger4 or Adv-Ptger4 ) ( n,o ) or EP2 (-Ptger2 ) ( P1p-Ptger2 or Adv-Ptger2 ) ( p,q ) or with AAV for a scrambled shRNA ( Plp-scrambled ) ( r ) ( n = 8 mice/group). Data are mean ± s.e.m. a , d , f , h , n , o , p , q , r 1-way or b , e , g , i , n , o , p , q , r 2-way ANOVA, Bonferroni correction. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs. Veh † P < 0.05, †† P < 0.01, ††† P < 0.001, †††† P < 0.0001 vs. PGE 2 /Veh, Control / PGE 2 . Source data are provided as a Source Data file.

Article Snippet: Human DRGs (#0062-HP-240, Gentaur), mouse DRGs, human sciatic nerve (#0062-HP-261, Gentaur), mouse paw tissue, and mouse sciatic nerve were incubated with different primary antibodies: EP2 (#ab167171, rabbit monoclonal, Abcam, 1:250 or #APR-064, rabbit polyclonal, Alomone Labs, 1:50), EP4 (#BS-8538R, rabbit polyclonal, Bioss, 1:200), S100B (#MA1-26621, mouse monoclonal, Invitrogen, 1:50 or #ab196175, rabbit monoclonal, Abcam, 1:50), Sox10 (#AF2864, goat polyclonal, R&D Systems, 1:100), NeuN (#MAB377, mouse monoclonal, Merck, 1:250 or #MAB377X, mouse monoclonal, Merck, 1:250), GFP (#A6455, rabbit polyclonal, Invitrogen, 1:500) and glutamine synthetase (#ab64613, mouse monoclonal, Abcam, 1:1000), myelin basic protein (MBP) (#ab133620, rabbit monoclonal, Abcam, 1:500), diluted in fresh blocking solution (PBS, pH 7.4, 5% normal goat serum (NGS) or normal donkey serum (NDS)).

Techniques: Injection, Quantitative RT-PCR, Expressing, Control, Infection, shRNA

Journal: Nature Communications

Article Title: Targeting prostaglandin E 2 receptor 2 in Schwann cells inhibits inflammatory pain but not inflammation

doi: 10.1038/s41467-025-63782-8

Figure Lengend Snippet: a Global cAMP formation in human Schwann cells (hSCs), after blue light stimulation (450 nm, 10 min) ( n = 121 cells, n = 3 independent experiments) or forskolin (FSK 100 μM) ( n = 78 cells, n = 3 independent experiments). b Schematic representation of Cre recombinase dependent expression of Beggiatoa photo-activable AC (bPAC) and mCherry separated by T2A self-cleaving peptide sequence. c Representative images of mCherry expression in S100B+ cells in mouse paw tissue after intraplantar (i.pl.) infection with AAV for selective expression of bPAC in SCs (scale bar: 20 μm) ( n = 4 subjects). d Illustration of bPAC activation after blue light stimulation and allodynia induced by blue light stimulation (pulsed 1 s/5 s for 10 min) in Plp-Cre or Control mice infected with AAV- bPAC ( Plp-bPAC ) ( n = 8 mice/group). e Representative images and cumulative data of membrane and intracellular localization of EP2 and EP4 in hSCs cells, with or without pre-incubation with unlabeled PGE 2 (10 µM, 30 min). Scale bar, 20 µm. ( n = 4 independent experiments). f Concentration-dependent global cAMP formation in hSCs induced by PGE 2 ( n = 8 replicates). g Membrane confined cAMP formation induced by PGE 2 (10 nM) in hSCs in the presence of PF-04418948 (PF,1 μM), BGC 20-1531 (BGC, 1 μM) or vehicle (Veh) (cells number: PGE 2 = 80, PF = 90, BGC = 88, n = 3 independent experiments). h Illustration of membrane tagged Lyn11-bPAC membrane confined cAMP formation and cAMP formation in hSCs after blue light stimulation ( n = 47 cells, n = 3 independent experiments) or FSK (100 μM) ( n = 46 cells, n = 3 independent experiments). i Allodynia induced by blue light stimulation in Plp-Cre or Control mice infected with AAV- Lyn11-bPAC ( Plp-Lyn11-bPAC ) ( n = 8 mice/group). j Concentration-dependent PKA activation induced by PGE 2 in hSCs ( n = 8 replicates). k Membrane confined PKA activation in hSCs induced by PGE 2 (10 nM) in the presence of PF (1 μM), BGC (1 μM) or Veh (cells number: PF = 64, BGC = 63, PGE 2 = 64, n = 3 independent experiments). Data are mean ± s.e.m. a , e , h ,2-tailed Student’s t test, g , k 1-way or d , i 2-way ANOVA, Bonferroni correction. AUC area under curve. **** P < 0.0001 vs. Control/blue light. Source data are provided as a Source Data file.

Article Snippet: Human DRGs (#0062-HP-240, Gentaur), mouse DRGs, human sciatic nerve (#0062-HP-261, Gentaur), mouse paw tissue, and mouse sciatic nerve were incubated with different primary antibodies: EP2 (#ab167171, rabbit monoclonal, Abcam, 1:250 or #APR-064, rabbit polyclonal, Alomone Labs, 1:50), EP4 (#BS-8538R, rabbit polyclonal, Bioss, 1:200), S100B (#MA1-26621, mouse monoclonal, Invitrogen, 1:50 or #ab196175, rabbit monoclonal, Abcam, 1:50), Sox10 (#AF2864, goat polyclonal, R&D Systems, 1:100), NeuN (#MAB377, mouse monoclonal, Merck, 1:250 or #MAB377X, mouse monoclonal, Merck, 1:250), GFP (#A6455, rabbit polyclonal, Invitrogen, 1:500) and glutamine synthetase (#ab64613, mouse monoclonal, Abcam, 1:1000), myelin basic protein (MBP) (#ab133620, rabbit monoclonal, Abcam, 1:500), diluted in fresh blocking solution (PBS, pH 7.4, 5% normal goat serum (NGS) or normal donkey serum (NDS)).

Techniques: Expressing, Sequencing, Infection, Activation Assay, Control, Membrane, Incubation, Concentration Assay