Journal: Reproduction (Cambridge, England)

Article Title: Increased expression of interleukin-1 receptor type 1 in active endometriotic lesions.

doi: 10.1530/rep.1.01121

Figure Lengend Snippet: Figure 1 Immunohistochemical analysis of IL-1RI expression in endometriotic tissue. (A) Note the intense brown immunostaining in the glands and cell aggregates throughout the stroma in the presence of a polyclonal goat anti-human IL-1RI antibody (a) and the absence of such staining in the presence of goat IgGs used at concentration equivalent to that of the primary antibody (b; negative control); scale bar, 30 mm. (B) HSCORE in red (nZ7), typical (nZ6), and white (nZ5) endometriotic lesions. G, gland, S, stroma; *, † and ‡significant difference with red lesions (P!0.05, !0.01, and !0.001 respectively) as analyzed by ANOVA and the Bonferroni’s test for multiple comparisons.

Article Snippet: Paraffin-embedded tissue sections were treated, then incubated for 30 min at room temperature with PBS/5% BSA (blocking solution) and incubated for 1 h at 37 8C with a polyclonal goat anti-human IL-1 receptor type I antibody (R&D Systems), at 15 mg/ml in PBS/BSA/Tween as described earlier (immunohistochemistry section).

Techniques: Immunohistochemical staining, Expressing, Immunostaining, Staining, Concentration Assay, Negative Control

Journal: Reproduction (Cambridge, England)

Article Title: Increased expression of interleukin-1 receptor type 1 in active endometriotic lesions.

doi: 10.1530/rep.1.01121

Figure Lengend Snippet: Figure 2 Dual-immunofluorescent staining of IL-1RI (A–C) and CD3 (D), CD68 (E), or vWF (F) in endometriotic tissue. Tissue sections were incubated with goat polyclonal anti- human IL-1RI antibody and with mouse monoclonal anti-CD3, mouse monoclonal CD68, or mouse monoclonal anti-human vWF antibody. Sections were then incubated simultaneously with fluorescein isothiocya- nate-conjugated donkey anti-goat antibody and rhodamine-conjugated sheep anti-mouse antibody to detect co-expression of IL-1RI with CD3, CD68, or vWF. Note the expression of IL-1RI (green) in CD3-, CD68-, and vWF- positive T-lymphocytes, macrophages, and endothelial cells respectively (red). Super- position of fluorescein (green) and rhodamine (red) signals clearly shows co-expression (yellow signal) of IL-1RI with CD3 (GZACD), CD68 (HZBCE), and vWF (IZCCF). Scale bars, 30 mm.

Article Snippet: Paraffin-embedded tissue sections were treated, then incubated for 30 min at room temperature with PBS/5% BSA (blocking solution) and incubated for 1 h at 37 8C with a polyclonal goat anti-human IL-1 receptor type I antibody (R&D Systems), at 15 mg/ml in PBS/BSA/Tween as described earlier (immunohistochemistry section).

Techniques: Staining, Incubation, Expressing

Journal: Journal of Neuroscience

Article Title: Nociceptors Are Interleukin-1 Sensors

doi: 10.1523/jneurosci.3795-08.2008

Figure Lengend Snippet: Figure 6. IL-1 modulates activity of DRG neurons via p38 MAPK. A, Increase in phosphorylated p38 MAP kinase (pp38) in culturedDRGneuronsafterIL-1(1ng/ml,10min)demonstratedbyimmunocytochemistryandWesternblot.B–E,Preincuba- tionofDRGneuronsfor40minwiththep38inhibitorSB203580(10M)preventedtheIL-1mediatedincreasesinexcitability. C,DosedependenteffectofSB203580onIL-1-mediateddecreaseofspikethreshold.(,10M;E,100nM;‚,10nM).Arrow indicates time of application of IL-1. Time point “-5” refers to data collected before application. Note no change in spike threshold after preincubation with 10 M SB203580 and significant decrease of threshold after preincubation with 10 nM SB203580.D,SB203580(10M)preventsIL-1-mediatedchangesinnumbersofspikesperstimulus(meanSEM)expressed as multiples of the rheobase for each recorded cell (n 10). E, Ramp (333 pA/s) activated trains of action potentials showed no changeinthresholdafterIL-1inpresenceofSB203580(10M),butstilldemonstratedreducedspikeadaptation.F,Theratioof themeanpeaktotalandTTX-rsodiumcurrentamplitude(IL-1,20min/control)(n10)showednochangeafterIL-1incells preincubated with 10 M SB203580 but increased significantly in presence of 10 nM SB203580. G, Quasi-steady-state current– voltagecurvesduringaslow(35mV/s)rampdemonstratingthatSB203580(10M)blockedtheIL-1-mediatedchangeinboth early(amplitudeVehicle14834pAvs15142pA,p0.23;currentactivationonsetVehicle538mVvs369mV,p 0.21;andpeakVehicle428mVvs4510mV,p0.11,n10)andlatecomponents(amplitudeVehicle11225pAvs 10512pA,p0.1;currentactivationonsetVehicle236mVvs244mV,p0.09andpeakVehicle125mVvs 165mV,p0.26,n10)ofresultinginwardcurrent.RecordingsforB–Eweremadeusingextracellularsolution4,those for F, solution 1, and those for G, solution 3 (Table 1).

Article Snippet: Double labeling was performed using rabbit anti IL-1R type I antibodies (R & D systems, 1:800), mouse anti peripherin antibodies (Millipore, 1:500), rabbit anti phospho-p38 MAP kinase (Cell Signaling, 1:600).

Techniques: Activity Assay, Control

Journal: Journal of Neuroscience

Article Title: Nociceptors Are Interleukin-1 Sensors

doi: 10.1523/jneurosci.3795-08.2008

Figure Lengend Snippet: Figure7. Activationofp38MAPKcontributestotheIL-1-mediatedbehavioraldecreaseinmechanicalthresholdandthermal responsiveness. A, Phosphorylation of p38 in peripherin-positive cutaneous sensory fibers after intraplantar IL-1 injection (1 ng). B, Pretreatment with the p38 inhibitor SB203580 (50 g) attenuated the effects of IL-1 on mechanical threshold and noxious heat response latency (mean SEM); *p 0.05.

Article Snippet: Double labeling was performed using rabbit anti IL-1R type I antibodies (R & D systems, 1:800), mouse anti peripherin antibodies (Millipore, 1:500), rabbit anti phospho-p38 MAP kinase (Cell Signaling, 1:600).

Techniques: Phospho-proteomics, Injection