Journal: Frontiers in endocrinology

Article Title: Excessive Iodine Promotes Pyroptosis of Thyroid Follicular Epithelial Cells in Hashimoto's Thyroiditis Through the ROS-NF-κB-NLRP3 Pathway.

doi: 10.3389/fendo.2019.00778

Figure Lengend Snippet: FIGURE 1 | Aberrant pyroptosis occurs in the thyroid tissues of HT patients and is induced by excessive iodine in vitro. (A,B) Gasdermin D (GSDMD) protein levels in the thyroid tissue of HT patients (n = 20) and controls (n = 10) were analyzed by immunoblots. “Control” indicates tissues from patients with nodular goiter of the thyroid. Representative immunoblotting results and quantification of GSDMD are shown. (C,D) Representative results of GSDMD immunohistochemical staining in HT tissues (n = 20) and control tissues (n = 10) are shown. Brown regions represent positive expression (original magnification, ×200; scale bars, 100 µm). (E–G) Nthy-ori3-1 cells were harvested after treatment with a gradient of concentrations of sodium iodide (NaI) for 24 h. The images presented are immunoblots probed for GSDMD (Hsp60 served as the loading control). All statistical results shown are representative of three replicates. (H) The cell viability of Nthy-ori 3-1 cells was assessed by CCK-8 assays after NaI treatment for 24 h. All statistical results shown are representative of three replicates. Significant differences and P-values were calculated by unpaired t-tests or one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Thyroid sections were blocked with 2% bovine serum albumin in PBS for 30min and then incubated with mouse anti-human GSDMD or mouse antihuman IL-1β antibodies (Santa Cruz, NJ, USA) at 4◦C overnight.

Techniques: In Vitro, Western Blot, Control, Immunohistochemical staining, Staining, Expressing, CCK-8 Assay

Journal: Frontiers in endocrinology

Article Title: Excessive Iodine Promotes Pyroptosis of Thyroid Follicular Epithelial Cells in Hashimoto's Thyroiditis Through the ROS-NF-κB-NLRP3 Pathway.

doi: 10.3389/fendo.2019.00778

Figure Lengend Snippet: FIGURE 3 | The NLRP3 inflammasome participates in excessive iodine-induced pyroptosis in TFCs. (A,B) The NLRP3 expression levels were measured by immunoblots when Nthy-ori 3-1 cells were treated with NaI with or without NAC (10 mM) or IKK-16 (2 µM) for 24 h. (C,D) Verification of the silencing efficiency of NLRP3 by siRNA in Nthy-ori3-1 cells is shown by immunoblots; NC indicates the negative control. (E,F) The protein levels of GSDMD were detected by immunoblots after transfection of siNLRP3 in NaI-treated Nthy-ori 3-1 cells. All statistical results shown are representative of three replicates. Significant differences and P-values were calculated by one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Thyroid sections were blocked with 2% bovine serum albumin in PBS for 30min and then incubated with mouse anti-human GSDMD or mouse antihuman IL-1β antibodies (Santa Cruz, NJ, USA) at 4◦C overnight.

Techniques: Expressing, Western Blot, Negative Control, Transfection

Journal: Frontiers in endocrinology

Article Title: Excessive Iodine Promotes Pyroptosis of Thyroid Follicular Epithelial Cells in Hashimoto's Thyroiditis Through the ROS-NF-κB-NLRP3 Pathway.

doi: 10.3389/fendo.2019.00778

Figure Lengend Snippet: FIGURE 5 | Proposed model of TFC pyroptosis in excessive iodine-promoted HT. Excessive iodine enters TFCs and induces the production of ROS, which activates NF-κB signaling and increases GSDMD-FL. In addition, the NLRP3 inflammasome is activated by NF-κB signaling, leading to GSDMD-FL cleavage into GSDMD-N, which triggers pore formation in the membrane and the release of mature IL-1β from cells, causing a sterile inflammatory response and further contributing to pyroptotic cell death and the subsequent promotion of HT.

Article Snippet: Thyroid sections were blocked with 2% bovine serum albumin in PBS for 30min and then incubated with mouse anti-human GSDMD or mouse antihuman IL-1β antibodies (Santa Cruz, NJ, USA) at 4◦C overnight.

Techniques: Membrane, Sterility

Journal: Kidney & blood pressure research

Article Title: Activation of TRPV1 Prevents Salt-Induced Kidney Damage and Hypertension After Renal Ischemia-Reperfusion Injury in Rats.

doi: 10.1159/000492412

Figure Lengend Snippet: Fig. 1. Effect of renal ischemia/reperfusion (I/R) and capsaicin pretreatment on the expression of renal TRPV1. Immunohistochemistry staining (a), representative Western blot (b) and quantification (c) of TRPV1 in renal tissue from rats in sham, I/R injury, I/R with pretreatment of a low dose of capsaicin (I/R+LCap), and I/R with pretreatment of a high dose of capsaicin (I/R+HCap) groups with HS diet for 4 weeks. Values are mean±SE (n=7 to 8). *P<0.05 compared with sham group; †P<0.05 compared with I/R group. Arrows indicate TRPV1 staining fibers. Scale bars, 50μm.

Article Snippet: USA), mouse anti-IL-1β antibody (1:200, sc-74138, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-CTGF antibody (1:200, sc-25440), rabbit anti-collagen I antibody (1:200, sc-8784), or rabbit anti-collagen IV antibody (1:200, sc-11360), at 4°C overnight, followed by incubation with HRP-donkey anti-rabbit IgG (1:10, 000, 711-035-152, Jackson ImmunoResearch Laboratories, West Grove, PA, USA) or HRP-donkey anti-mouse IgG (1:10, 000, 711-035-151, Jackson ImmunoResearch Laboratories) for 2h at room temperature.

Techniques: Expressing, Immunohistochemistry, Staining, Western Blot

Journal: Kidney & blood pressure research

Article Title: Activation of TRPV1 Prevents Salt-Induced Kidney Damage and Hypertension After Renal Ischemia-Reperfusion Injury in Rats.

doi: 10.1159/000492412

Figure Lengend Snippet: Fig. 3. Effect of I/R, capsaicin pretreatment and high salt loading on renal function. 24-hour ratio of urine/ water intake m e a s u r e d once a week before and after I/R (a), and creatinine clearance (b) and plasma urea (c) and creatinine (d) measured at 1 day after I/R, 5-week after I/R, and 4-week after high salt loading in rats of sham, I/R injury, I/R+LCap, and I/R+HCap groups. Values are mean±SE (n=6-8). *P<0.05 compared with the sham group; †P<0.05 compared with the I/R group.

Article Snippet: USA), mouse anti-IL-1β antibody (1:200, sc-74138, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-CTGF antibody (1:200, sc-25440), rabbit anti-collagen I antibody (1:200, sc-8784), or rabbit anti-collagen IV antibody (1:200, sc-11360), at 4°C overnight, followed by incubation with HRP-donkey anti-rabbit IgG (1:10, 000, 711-035-152, Jackson ImmunoResearch Laboratories, West Grove, PA, USA) or HRP-donkey anti-mouse IgG (1:10, 000, 711-035-151, Jackson ImmunoResearch Laboratories) for 2h at room temperature.

Techniques: Clinical Proteomics

Journal: Kidney & blood pressure research

Article Title: Activation of TRPV1 Prevents Salt-Induced Kidney Damage and Hypertension After Renal Ischemia-Reperfusion Injury in Rats.

doi: 10.1159/000492412

Figure Lengend Snippet: Fig. 4. Effect of I/R, capsaicin pretreatment and high salt loading on renal inflammation. Western blotting for the expression of TNF-α (a, b) and IL-1β (c, d) in the kidney from rats in sham, I/R injury, I/R+LCap, and I/R+HCap groups with HS diet for 4 weeks. Values are mean±SE (n=5). *P<0.05 compared with the sham group; †P<0.05 compared with the I/R group.

Article Snippet: USA), mouse anti-IL-1β antibody (1:200, sc-74138, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-CTGF antibody (1:200, sc-25440), rabbit anti-collagen I antibody (1:200, sc-8784), or rabbit anti-collagen IV antibody (1:200, sc-11360), at 4°C overnight, followed by incubation with HRP-donkey anti-rabbit IgG (1:10, 000, 711-035-152, Jackson ImmunoResearch Laboratories, West Grove, PA, USA) or HRP-donkey anti-mouse IgG (1:10, 000, 711-035-151, Jackson ImmunoResearch Laboratories) for 2h at room temperature.

Techniques: Western Blot, Expressing

Journal: Nature Communications

Article Title: Schwann cell C5aR1 co-opts inflammasome NLRP1 to sustain pain in a mouse model of endometriosis

doi: 10.1038/s41467-024-54486-6

Figure Lengend Snippet: a Time-dependent periorbital (PMA), hind paw (HMA) and abdominal (AMA) mechanical allodynia in endometriotic (endo) or Sham C57BL/6 J female (B6) mice. Proteome profiler array ( b ) and single analyte C5a assay ( c ) in plasma samples from endo and Sham B6 mice. d C5 assay in plasma samples from endo patients ( n = 19) or healthy subjects ( n = 15) (control). e Time-dependent C5a levels in plasma samples from endo and Sham B6 mice. f and g Timeline of the DF2593A (DF25) treatment schedule and time-dependent PMA, HMA, and AMA in endo B6 mice after intragastric (i.g.) administration of DF25 (1 mg/kg) or vehicle (Veh) and Sham mice. ( n = 8 mice per group). Data are mean ± s.e.m. a , e , f , g 2-way ANOVA, Bonferroni correction; c , d 2- tailed Student’s t test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs. Sham ## P < 0.01, #### P < 0.0001 vs. Endo+Veh DF25. Source data are provided as a Source Data file.

Article Snippet: After blocking using normal donkey serum (NDS, 5%) for 1 h sciatic and trigeminal nerve tissue were incubated with the following primary antibodies: F4/80 (#MA516624, rat monoclonal (Cl:A3-1), Thermo Fisher Scientific, 1:50, RRID:AB_253820), C5aR1 (#orb393229, rabbit polyclonal, Biorbyt Ltd, 1:100), C5aR1 (#MCA2456GA, rat monoclonal, Bio-Rad, 1:100, RRID:AB_770091), NLRP3 (#MA5-32255, rabbit monoclonal, Thermo Fisher Scientific, 1:100, RRID:AB_2809541), NLRP1 (#12256-1-AP, rabbit polyclonal, Proteintech, 1:50, RRID:AB_2298504) and IL-1β (#NB600-633, rabbit polyclonal, Novus Biological, 1:100, RRID:AB_10001060) for 1 h at room temperature and O/N at 4 °C with SOX10 (#AF2864, goat polyclonal, R&D Systems, 1:200, RRID:AB_442208).

Techniques: Clinical Proteomics, Control

Journal: Nature Communications

Article Title: Schwann cell C5aR1 co-opts inflammasome NLRP1 to sustain pain in a mouse model of endometriosis

doi: 10.1038/s41467-024-54486-6

Figure Lengend Snippet: a Representative images and cumulative data of the time-dependent increase of F4/80 + cells in sciatic and trigeminal nerve in endometriotic (endo) or Sham C57BL/6 J female (B6) mice. ( n = 4 independent experiments). b Time-dependent C5a levels in sciatic and trigeminal nerve homogenates from endo and Sham B6 mice. ( n = 6 independent experiments). c Timeline of the AP20187 (AP) treatment schedule and time-dependent periorbital (PMA), hind paw (HMA) and abdominal (AMA) mechanical allodynia in endo and Sham MaFIA mice after intraperitoneal (i.p.) injection of AP or vehicle (Veh). ( n = 8 mice per group). d Representative images and cumulative data of F4/80 + cells in sciatic and trigeminal nerve of endo B6 mice after AP or Veh and in Sham mice (treatment schedule as in b ). ( n = 4 independent experiments). (Scale bar, 50 μm, dashed lines, perineurium ). Data are mean ± s.e.m. a, b, d 1-way ANOVA, c , 2-way ANOVA, Bonferroni correction; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs. Sham, ### # P < 0.0001 vs. Endo-MaFIA+ Veh AP. Source data are provided as a Source Data file.

Article Snippet: After blocking using normal donkey serum (NDS, 5%) for 1 h sciatic and trigeminal nerve tissue were incubated with the following primary antibodies: F4/80 (#MA516624, rat monoclonal (Cl:A3-1), Thermo Fisher Scientific, 1:50, RRID:AB_253820), C5aR1 (#orb393229, rabbit polyclonal, Biorbyt Ltd, 1:100), C5aR1 (#MCA2456GA, rat monoclonal, Bio-Rad, 1:100, RRID:AB_770091), NLRP3 (#MA5-32255, rabbit monoclonal, Thermo Fisher Scientific, 1:100, RRID:AB_2809541), NLRP1 (#12256-1-AP, rabbit polyclonal, Proteintech, 1:50, RRID:AB_2298504) and IL-1β (#NB600-633, rabbit polyclonal, Novus Biological, 1:100, RRID:AB_10001060) for 1 h at room temperature and O/N at 4 °C with SOX10 (#AF2864, goat polyclonal, R&D Systems, 1:200, RRID:AB_442208).

Techniques: Injection

Journal: Nature Communications

Article Title: Schwann cell C5aR1 co-opts inflammasome NLRP1 to sustain pain in a mouse model of endometriosis

doi: 10.1038/s41467-024-54486-6

Figure Lengend Snippet: Representative images of SOX10 and C5aR1 (#orb393229, rabbit polyclonal, Biorbyt Ltd, 1:100) co-expression in sciatic ( a ) and trigeminal ( b ) nerve tissue from C57BL/6 J female (B6) mice, ( n = 4 independent experiments) (Scale bar, 50 μm). (c) C5aR1 mRNA relative expression in primary mouse and human Schwann cells ( n = 3 independent experiments). d Schematic representation of AAV-(loxP-shRNA)- C5aR1 vector pre- and post-Cre switch. e Representative images and cumulative data (Rcoloc) of SOX10 and C5aR1 (#orb393229, rabbit polyclonal, Biorbyt Ltd, 1:100) co-expression in sciatic and trigeminal nerve tissue in Plp -AAV- C5aR1 and Control mice ( n = 4 independent experiments) (Scale bar, 50 μm). f Time-dependent periorbital (PMA), hind paw (HMA), and abdominal (AMA) mechanical allodynia in endometriotic (endo) or Sham Plp -AAV- C5aR1 and Control mice. ( n = 8 mice per group). g Representative images and cumulative data of F4/80 + cells in sciatic and trigeminal nerve tissue in endo or Sham Plp -AAV- C5aR1 and Control mice. ( n = 4 independent experiments) (Scale bar, 50 μm, dashed lines, perineurium ). h Representative images and cumulative data of transmigrated macrophages after stimulation of human and mouse Schwann cells with C5a or vehicle (Veh) and in the presence of DF2593A (DF25) or Veh ( n = 6 independent experiments). Data are mean ± s.e.m. c , Student’s t-test, f , 2-way, g, h 1-way ANOVA, Bonferroni correction; **** P < 0.0001 vs. Sham/Control, #### P < 0.0001 vs. Endo/Control. Source data are provided as a Source Data file.

Article Snippet: After blocking using normal donkey serum (NDS, 5%) for 1 h sciatic and trigeminal nerve tissue were incubated with the following primary antibodies: F4/80 (#MA516624, rat monoclonal (Cl:A3-1), Thermo Fisher Scientific, 1:50, RRID:AB_253820), C5aR1 (#orb393229, rabbit polyclonal, Biorbyt Ltd, 1:100), C5aR1 (#MCA2456GA, rat monoclonal, Bio-Rad, 1:100, RRID:AB_770091), NLRP3 (#MA5-32255, rabbit monoclonal, Thermo Fisher Scientific, 1:100, RRID:AB_2809541), NLRP1 (#12256-1-AP, rabbit polyclonal, Proteintech, 1:50, RRID:AB_2298504) and IL-1β (#NB600-633, rabbit polyclonal, Novus Biological, 1:100, RRID:AB_10001060) for 1 h at room temperature and O/N at 4 °C with SOX10 (#AF2864, goat polyclonal, R&D Systems, 1:200, RRID:AB_442208).

Techniques: Expressing, shRNA, Plasmid Preparation, Control

Journal: Nature Communications

Article Title: Schwann cell C5aR1 co-opts inflammasome NLRP1 to sustain pain in a mouse model of endometriosis

doi: 10.1038/s41467-024-54486-6

Figure Lengend Snippet: a IL-1β assay in human Schwann cell conditioned medium after stimulation with C5a or vehicle (Veh) and in the presence of DF2593A (DF25) or Veh ( n = 6 independent experiments). b Time-dependent C5a levels in plasma samples from endometriotic (endo) or Sham C57BL/6 J female (B6) mice. ( n = 8 independent experiments). c IL-1β assay in plasma samples from endo patients ( n = 16) or healthy subjects ( n = 15) (control). d Typical traces and cumulative data of SCAT1 V/C ratio changes in human Schwann cells stimulated with C5a or Veh and in the presence of DF25or Ac-YVAD (AUC, area under the curve) (cell numbers: C5a = 20, Veh=16, DF25 = 16 and Ac-YVAD = 20; n = 3 independent experiments) e Time-dependent periorbital (PMA), hind paw (HMA) and abdominal (AMA) mechanical allodynia in endo or Sham Plp- AAV- Nlrp1 and Control mice ( n = 8 mice per group). f IL-1β assay in sciatic and trigeminal nerve tissue homogenates in endo or Sham Plp- AAV- Nlrp1 and Control mice ( n = 6 independent experiments). g Representative images and cumulative data of F4/80 + cells in sciatic and trigeminal nerves in endo or Sham Plp- AAV- Nlrp1 and Control mice ( n = 4 independent experiments). h Time-dependent PMA, HMA, and AMA in endo or Sham Plp- AAV- IL-1β and Control mice ( n = 8 mice per group). i Il-1β assay in sciatic and trigeminal nerve tissue homogenates in endo or Sham Plp- AAV- Il-1β and Control mice. j Representative images and cumulative data of F4/80 + cells in sciatic and periorbital nerve in endo or Sham Plp- AAV- Il-1β and Control mice (Scale bar, 50 μm, dashed lines, perineurium ) ( n = 8 mice per group). Data are mean ± s.e.m. a, c, d, f, g 1-way ANOVA, b, e, h , 2-way ANOVA, Bonferroni correction; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs. Sham, # P < 0.05, ## P < 0.01, ### P < 0.001, #### P < 0.0001 vs. Endo/Control. Source data are provided as a Source Data file.

Article Snippet: After blocking using normal donkey serum (NDS, 5%) for 1 h sciatic and trigeminal nerve tissue were incubated with the following primary antibodies: F4/80 (#MA516624, rat monoclonal (Cl:A3-1), Thermo Fisher Scientific, 1:50, RRID:AB_253820), C5aR1 (#orb393229, rabbit polyclonal, Biorbyt Ltd, 1:100), C5aR1 (#MCA2456GA, rat monoclonal, Bio-Rad, 1:100, RRID:AB_770091), NLRP3 (#MA5-32255, rabbit monoclonal, Thermo Fisher Scientific, 1:100, RRID:AB_2809541), NLRP1 (#12256-1-AP, rabbit polyclonal, Proteintech, 1:50, RRID:AB_2298504) and IL-1β (#NB600-633, rabbit polyclonal, Novus Biological, 1:100, RRID:AB_10001060) for 1 h at room temperature and O/N at 4 °C with SOX10 (#AF2864, goat polyclonal, R&D Systems, 1:200, RRID:AB_442208).

Techniques: Clinical Proteomics, Control

Journal: Nature Communications

Article Title: Schwann cell C5aR1 co-opts inflammasome NLRP1 to sustain pain in a mouse model of endometriosis

doi: 10.1038/s41467-024-54486-6

Figure Lengend Snippet: a Time-dependent periorbital (PMA), hind paw (HMA) and abdominal (AMA) mechanical allodynia in endometriotic (endo) or Sham Plp-Trpa1 and Control mice ( n = 8 mice per group). Representative images and cumulative data of ( b ) F4/80 + cells and ( c ) 4-HNE staining in sciatic and trigeminal nerve in endo or Sham Plp-Trpa1 and Control mice (Scale bar, 50 μm, dashed lines, perineurium ) ( n = 4 and n = 5 independent experiments). d Time-dependent PMA, HMA and AMA in endometriotic (endo) or Sham Adv-Trpa1 and Control mice ( n = 8 mice per group). Representative images and cumulative data of ( e ) F4/80 + cells and ( f ) 4-HNE staining in sciatic and trigeminal nerve in endo or Sham Adv-Trpa1 and Control mice (Scale bar, 50 μm, dashed lines, perineurium ) ( n = 4 independent experiments). Data are mean ± s.e.m. a, d 2-way ANOVA, b,c,e,f 1-way ANOVA, Bonferroni correction; * P < 0.05, *** P < 0.001, **** P < 0.0001 vs. Sham, # P < 0.05, # # P < 0.01, ### P < 0.001, #### P < 0.0001 vs. Endo/Control. Source data are provided as a Source Data file.

Article Snippet: After blocking using normal donkey serum (NDS, 5%) for 1 h sciatic and trigeminal nerve tissue were incubated with the following primary antibodies: F4/80 (#MA516624, rat monoclonal (Cl:A3-1), Thermo Fisher Scientific, 1:50, RRID:AB_253820), C5aR1 (#orb393229, rabbit polyclonal, Biorbyt Ltd, 1:100), C5aR1 (#MCA2456GA, rat monoclonal, Bio-Rad, 1:100, RRID:AB_770091), NLRP3 (#MA5-32255, rabbit monoclonal, Thermo Fisher Scientific, 1:100, RRID:AB_2809541), NLRP1 (#12256-1-AP, rabbit polyclonal, Proteintech, 1:50, RRID:AB_2298504) and IL-1β (#NB600-633, rabbit polyclonal, Novus Biological, 1:100, RRID:AB_10001060) for 1 h at room temperature and O/N at 4 °C with SOX10 (#AF2864, goat polyclonal, R&D Systems, 1:200, RRID:AB_442208).

Techniques: Control, Staining

Journal:

Article Title: Cytokine response to vitamin E supplementation is dependent on pre-supplementation cytokine levels

doi:

Figure Lengend Snippet: Description of SNPs, primers and probes

Article Snippet: IL-1β was detected using mouse anti-human IL-1β MAb and biotinylated anti-human IL-1β Ab (R&D Systems, Minneapolis, MN).

Techniques:

Journal:

Article Title: Cytokine response to vitamin E supplementation is dependent on pre-supplementation cytokine levels

doi:

Figure Lengend Snippet: Genotype frequencies

Article Snippet: IL-1β was detected using mouse anti-human IL-1β MAb and biotinylated anti-human IL-1β Ab (R&D Systems, Minneapolis, MN).

Techniques:

Journal:

Article Title: Cytokine response to vitamin E supplementation is dependent on pre-supplementation cytokine levels

doi:

Figure Lengend Snippet: The effect of vitamin E supplementation on cytokine production depends on baseline cytokine production. Cytokine production was measured from whole blood at the beginning and end of a one year vitamin E supplementation in the elderly. TNF-α, IL-1β, and IL-6 was measured from whole blood elicited for 24 hours with lipopolysacchride (LPS; 1.0 µg/mL). IFN-γ was measured from whole blood elicited for 48 hours with phytohemagluttinin (PHA; 20 µg/mL) or concalavinA (ConA; 40 µg/mL). Cytokines production was corrected for the number of monocytes and lymphocytes in the blood (pg/ 106 lymphocyte and monocyte). P values (P) for the interaction are unadjusted.

Article Snippet: IL-1β was detected using mouse anti-human IL-1β MAb and biotinylated anti-human IL-1β Ab (R&D Systems, Minneapolis, MN).

Techniques: