mouse anti saa1 a2 Search Results


goat anti mouse saa1 antibody  (R&D Systems)
99
R&D Systems goat anti mouse saa1 antibody
Goat Anti Mouse Saa1 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/goat anti mouse saa1 antibody/product/R&D Systems
Average 99 stars, based on 1 article reviews
goat anti mouse saa1 antibody - by Bioz Stars, 2026-03
99/100 stars
  Buy from Supplier
biotinylated anti mouse saa1 antibody  (R&D Systems)
91
R&D Systems biotinylated anti mouse saa1 antibody
Biotinylated Anti Mouse Saa1 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/biotinylated anti mouse saa1 antibody/product/R&D Systems
Average 91 stars, based on 1 article reviews
biotinylated anti mouse saa1 antibody - by Bioz Stars, 2026-03
91/100 stars
  Buy from Supplier
mouse anti human saa1 antibody  (R&D Systems)
92
R&D Systems mouse anti human saa1 antibody
(a) Model of allergen-induced airway hyperresponsiveness (AHR) for wild-type (WT) and Saa–/– mice (both C57BL/6 background). Mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 μg of HDM extract. Airway measurements were performed 72 h after the last allergen challenge (used in Fig. 1a-​-jj and Extended Data Fig. 2 and ​and3).3). (b) For <t>SAA1</t> antibody blockade, we used an established mouse model of allergen-induced AHR sensitizing WT BALB/cJ mice i.t. on day 0 and 14 with 100 µg of HDM extract + isotype control, or HDM + αSAAab. Airway measurements and tissue harvests were performed 72 h after the last allergen challenge (used in Extended Data Fig. 4). (c) In short-term exposure protocols WT and Saa–/– mice (both C57BL/6 background) received a single HDM challenge (100 μg) were sacrificed 16 h later (used in Fig. 2a-​-d).d). (d) In short-term exposure protocols BALB/cJ mice received a single HDM challenge (100 μg)+ isotype control, HDM challenge + HDL (200μg) and isotype control, or HDM + αSAAab and were sacrificed 24 h later (used in Fig. 2e). Contol mice received either PBS + isotype or PBS + HDL and isotype control. (e) For overexpression of SAA1 in vivo, mice were injected 20 µg of DNA complexed to polyethylenimine at day 0, exposed to PBS or HDM 48 h later and ILC2s as well as BAL cytokines measurements were performed on day 3 (used Fig. 2f). (f) WT and Saa–/– mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 μg with extracts from the parasitic worm Schistosoma mansoni (a Puerto Rican isolate). Tissues were harvested 72 h after the last allergen challenge (used in Fig. 4). (g) For FPR2 blockade (WRW4, 2 mg/kg), WT BALB/cJ mice were sensitized and challenged i.t. on day 0 and 14 with 100 μg of HDM extract. Airway measurements were performed 72 h after the last allergen challenge (used in Fig. 7a-​-f).f). (h) In short term exposure experiments, BALB/cJ mice received a single HDM challenge (100 μg) or HDM + WRW4 and were sacrificed 24 h later (used in Fig. 7g and ​andi).i). (i) Model of Alternaria alternata (Alt a )-induced airway inflammation for WT and Saa–/– mice. Mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 µg of Alt a extract. Tissues were harvested 72 h after the last allergen challenge (used in Extended Data Fig. 9f-​-jj).
Mouse Anti Human Saa1 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse anti human saa1 antibody/product/R&D Systems
Average 92 stars, based on 1 article reviews
mouse anti human saa1 antibody - by Bioz Stars, 2026-03
92/100 stars
  Buy from Supplier
goat-anti-mouse  (Millipore)
90
Millipore goat-anti-mouse
(a) Model of allergen-induced airway hyperresponsiveness (AHR) for wild-type (WT) and Saa–/– mice (both C57BL/6 background). Mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 μg of HDM extract. Airway measurements were performed 72 h after the last allergen challenge (used in Fig. 1a-​-jj and Extended Data Fig. 2 and ​and3).3). (b) For <t>SAA1</t> antibody blockade, we used an established mouse model of allergen-induced AHR sensitizing WT BALB/cJ mice i.t. on day 0 and 14 with 100 µg of HDM extract + isotype control, or HDM + αSAAab. Airway measurements and tissue harvests were performed 72 h after the last allergen challenge (used in Extended Data Fig. 4). (c) In short-term exposure protocols WT and Saa–/– mice (both C57BL/6 background) received a single HDM challenge (100 μg) were sacrificed 16 h later (used in Fig. 2a-​-d).d). (d) In short-term exposure protocols BALB/cJ mice received a single HDM challenge (100 μg)+ isotype control, HDM challenge + HDL (200μg) and isotype control, or HDM + αSAAab and were sacrificed 24 h later (used in Fig. 2e). Contol mice received either PBS + isotype or PBS + HDL and isotype control. (e) For overexpression of SAA1 in vivo, mice were injected 20 µg of DNA complexed to polyethylenimine at day 0, exposed to PBS or HDM 48 h later and ILC2s as well as BAL cytokines measurements were performed on day 3 (used Fig. 2f). (f) WT and Saa–/– mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 μg with extracts from the parasitic worm Schistosoma mansoni (a Puerto Rican isolate). Tissues were harvested 72 h after the last allergen challenge (used in Fig. 4). (g) For FPR2 blockade (WRW4, 2 mg/kg), WT BALB/cJ mice were sensitized and challenged i.t. on day 0 and 14 with 100 μg of HDM extract. Airway measurements were performed 72 h after the last allergen challenge (used in Fig. 7a-​-f).f). (h) In short term exposure experiments, BALB/cJ mice received a single HDM challenge (100 μg) or HDM + WRW4 and were sacrificed 24 h later (used in Fig. 7g and ​andi).i). (i) Model of Alternaria alternata (Alt a )-induced airway inflammation for WT and Saa–/– mice. Mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 µg of Alt a extract. Tissues were harvested 72 h after the last allergen challenge (used in Extended Data Fig. 9f-​-jj).
Goat Anti Mouse, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/goat-anti-mouse/product/Millipore
Average 90 stars, based on 1 article reviews
goat-anti-mouse - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier
mouse anti saa1 a2  (R&D Systems)
92
R&D Systems mouse anti saa1 a2
(a) Model of allergen-induced airway hyperresponsiveness (AHR) for wild-type (WT) and Saa–/– mice (both C57BL/6 background). Mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 μg of HDM extract. Airway measurements were performed 72 h after the last allergen challenge (used in Fig. 1a-​-jj and Extended Data Fig. 2 and ​and3).3). (b) For <t>SAA1</t> antibody blockade, we used an established mouse model of allergen-induced AHR sensitizing WT BALB/cJ mice i.t. on day 0 and 14 with 100 µg of HDM extract + isotype control, or HDM + αSAAab. Airway measurements and tissue harvests were performed 72 h after the last allergen challenge (used in Extended Data Fig. 4). (c) In short-term exposure protocols WT and Saa–/– mice (both C57BL/6 background) received a single HDM challenge (100 μg) were sacrificed 16 h later (used in Fig. 2a-​-d).d). (d) In short-term exposure protocols BALB/cJ mice received a single HDM challenge (100 μg)+ isotype control, HDM challenge + HDL (200μg) and isotype control, or HDM + αSAAab and were sacrificed 24 h later (used in Fig. 2e). Contol mice received either PBS + isotype or PBS + HDL and isotype control. (e) For overexpression of SAA1 in vivo, mice were injected 20 µg of DNA complexed to polyethylenimine at day 0, exposed to PBS or HDM 48 h later and ILC2s as well as BAL cytokines measurements were performed on day 3 (used Fig. 2f). (f) WT and Saa–/– mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 μg with extracts from the parasitic worm Schistosoma mansoni (a Puerto Rican isolate). Tissues were harvested 72 h after the last allergen challenge (used in Fig. 4). (g) For FPR2 blockade (WRW4, 2 mg/kg), WT BALB/cJ mice were sensitized and challenged i.t. on day 0 and 14 with 100 μg of HDM extract. Airway measurements were performed 72 h after the last allergen challenge (used in Fig. 7a-​-f).f). (h) In short term exposure experiments, BALB/cJ mice received a single HDM challenge (100 μg) or HDM + WRW4 and were sacrificed 24 h later (used in Fig. 7g and ​andi).i). (i) Model of Alternaria alternata (Alt a )-induced airway inflammation for WT and Saa–/– mice. Mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 µg of Alt a extract. Tissues were harvested 72 h after the last allergen challenge (used in Extended Data Fig. 9f-​-jj).
Mouse Anti Saa1 A2, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse anti saa1 a2/product/R&D Systems
Average 92 stars, based on 1 article reviews
mouse anti saa1 a2 - by Bioz Stars, 2026-03
92/100 stars
  Buy from Supplier
diaclone research b c50 mouse anti saa1 a2 r d systems 924903 goat  (R&D Systems)
92
R&D Systems diaclone research b c50 mouse anti saa1 a2 r d systems 924903 goat
(a) Model of allergen-induced airway hyperresponsiveness (AHR) for wild-type (WT) and Saa–/– mice (both C57BL/6 background). Mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 μg of HDM extract. Airway measurements were performed 72 h after the last allergen challenge (used in Fig. 1a-​-jj and Extended Data Fig. 2 and ​and3).3). (b) For <t>SAA1</t> antibody blockade, we used an established mouse model of allergen-induced AHR sensitizing WT BALB/cJ mice i.t. on day 0 and 14 with 100 µg of HDM extract + isotype control, or HDM + αSAAab. Airway measurements and tissue harvests were performed 72 h after the last allergen challenge (used in Extended Data Fig. 4). (c) In short-term exposure protocols WT and Saa–/– mice (both C57BL/6 background) received a single HDM challenge (100 μg) were sacrificed 16 h later (used in Fig. 2a-​-d).d). (d) In short-term exposure protocols BALB/cJ mice received a single HDM challenge (100 μg)+ isotype control, HDM challenge + HDL (200μg) and isotype control, or HDM + αSAAab and were sacrificed 24 h later (used in Fig. 2e). Contol mice received either PBS + isotype or PBS + HDL and isotype control. (e) For overexpression of SAA1 in vivo, mice were injected 20 µg of DNA complexed to polyethylenimine at day 0, exposed to PBS or HDM 48 h later and ILC2s as well as BAL cytokines measurements were performed on day 3 (used Fig. 2f). (f) WT and Saa–/– mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 μg with extracts from the parasitic worm Schistosoma mansoni (a Puerto Rican isolate). Tissues were harvested 72 h after the last allergen challenge (used in Fig. 4). (g) For FPR2 blockade (WRW4, 2 mg/kg), WT BALB/cJ mice were sensitized and challenged i.t. on day 0 and 14 with 100 μg of HDM extract. Airway measurements were performed 72 h after the last allergen challenge (used in Fig. 7a-​-f).f). (h) In short term exposure experiments, BALB/cJ mice received a single HDM challenge (100 μg) or HDM + WRW4 and were sacrificed 24 h later (used in Fig. 7g and ​andi).i). (i) Model of Alternaria alternata (Alt a )-induced airway inflammation for WT and Saa–/– mice. Mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 µg of Alt a extract. Tissues were harvested 72 h after the last allergen challenge (used in Extended Data Fig. 9f-​-jj).
Diaclone Research B C50 Mouse Anti Saa1 A2 R D Systems 924903 Goat, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/diaclone research b c50 mouse anti saa1 a2 r d systems 924903 goat/product/R&D Systems
Average 92 stars, based on 1 article reviews
diaclone research b c50 mouse anti saa1 a2 r d systems 924903 goat - by Bioz Stars, 2026-03
92/100 stars
  Buy from Supplier
against mouse saa 1  (R&D Systems)
86
R&D Systems against mouse saa 1
(a) Model of allergen-induced airway hyperresponsiveness (AHR) for wild-type (WT) and Saa–/– mice (both C57BL/6 background). Mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 μg of HDM extract. Airway measurements were performed 72 h after the last allergen challenge (used in Fig. 1a-​-jj and Extended Data Fig. 2 and ​and3).3). (b) For <t>SAA1</t> antibody blockade, we used an established mouse model of allergen-induced AHR sensitizing WT BALB/cJ mice i.t. on day 0 and 14 with 100 µg of HDM extract + isotype control, or HDM + αSAAab. Airway measurements and tissue harvests were performed 72 h after the last allergen challenge (used in Extended Data Fig. 4). (c) In short-term exposure protocols WT and Saa–/– mice (both C57BL/6 background) received a single HDM challenge (100 μg) were sacrificed 16 h later (used in Fig. 2a-​-d).d). (d) In short-term exposure protocols BALB/cJ mice received a single HDM challenge (100 μg)+ isotype control, HDM challenge + HDL (200μg) and isotype control, or HDM + αSAAab and were sacrificed 24 h later (used in Fig. 2e). Contol mice received either PBS + isotype or PBS + HDL and isotype control. (e) For overexpression of SAA1 in vivo, mice were injected 20 µg of DNA complexed to polyethylenimine at day 0, exposed to PBS or HDM 48 h later and ILC2s as well as BAL cytokines measurements were performed on day 3 (used Fig. 2f). (f) WT and Saa–/– mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 μg with extracts from the parasitic worm Schistosoma mansoni (a Puerto Rican isolate). Tissues were harvested 72 h after the last allergen challenge (used in Fig. 4). (g) For FPR2 blockade (WRW4, 2 mg/kg), WT BALB/cJ mice were sensitized and challenged i.t. on day 0 and 14 with 100 μg of HDM extract. Airway measurements were performed 72 h after the last allergen challenge (used in Fig. 7a-​-f).f). (h) In short term exposure experiments, BALB/cJ mice received a single HDM challenge (100 μg) or HDM + WRW4 and were sacrificed 24 h later (used in Fig. 7g and ​andi).i). (i) Model of Alternaria alternata (Alt a )-induced airway inflammation for WT and Saa–/– mice. Mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 µg of Alt a extract. Tissues were harvested 72 h after the last allergen challenge (used in Extended Data Fig. 9f-​-jj).
Against Mouse Saa 1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/against mouse saa 1/product/R&D Systems
Average 86 stars, based on 1 article reviews
against mouse saa 1 - by Bioz Stars, 2026-03
86/100 stars
  Buy from Supplier
primary antibody against saa1  (Proteintech)
93
Proteintech primary antibody against saa1
The schematic diagram of the experiment design. CYP2D6 cytochrome P450 2D6, AIH autoimmune hepatitis, DEPs differentially expressed proteins, iTRAQ isobaric tags for relative and absolute quantitation, <t>SAA1</t> serum amyloid A 1, GO gene ontology
Primary Antibody Against Saa1, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/primary antibody against saa1/product/Proteintech
Average 93 stars, based on 1 article reviews
primary antibody against saa1 - by Bioz Stars, 2026-03
93/100 stars
  Buy from Supplier
mouse-anti-human ip-10 mab b-c55  (Diaclone)
90
Diaclone mouse-anti-human ip-10 mab b-c55
The schematic diagram of the experiment design. CYP2D6 cytochrome P450 2D6, AIH autoimmune hepatitis, DEPs differentially expressed proteins, iTRAQ isobaric tags for relative and absolute quantitation, <t>SAA1</t> serum amyloid A 1, GO gene ontology
Mouse Anti Human Ip 10 Mab B C55, supplied by Diaclone, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse-anti-human ip-10 mab b-c55/product/Diaclone
Average 90 stars, based on 1 article reviews
mouse-anti-human ip-10 mab b-c55 - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier
luminescent up-converting reporter particles ucps nayf4:yb 3+,er 3  (Intelligent Material Solutions)
90
Intelligent Material Solutions luminescent up-converting reporter particles ucps nayf4:yb 3+,er 3
The schematic diagram of the experiment design. CYP2D6 cytochrome P450 2D6, AIH autoimmune hepatitis, DEPs differentially expressed proteins, iTRAQ isobaric tags for relative and absolute quantitation, <t>SAA1</t> serum amyloid A 1, GO gene ontology
Luminescent Up Converting Reporter Particles Ucps Nayf4:Yb 3+,Er 3, supplied by Intelligent Material Solutions, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/luminescent up-converting reporter particles ucps nayf4:yb 3+,er 3/product/Intelligent Material Solutions
Average 90 stars, based on 1 article reviews
luminescent up-converting reporter particles ucps nayf4:yb 3+,er 3 - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier

Image Search Results


(a) Model of allergen-induced airway hyperresponsiveness (AHR) for wild-type (WT) and Saa–/– mice (both C57BL/6 background). Mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 μg of HDM extract. Airway measurements were performed 72 h after the last allergen challenge (used in Fig. 1a-​-jj and Extended Data Fig. 2 and ​and3).3). (b) For SAA1 antibody blockade, we used an established mouse model of allergen-induced AHR sensitizing WT BALB/cJ mice i.t. on day 0 and 14 with 100 µg of HDM extract + isotype control, or HDM + αSAAab. Airway measurements and tissue harvests were performed 72 h after the last allergen challenge (used in Extended Data Fig. 4). (c) In short-term exposure protocols WT and Saa–/– mice (both C57BL/6 background) received a single HDM challenge (100 μg) were sacrificed 16 h later (used in Fig. 2a-​-d).d). (d) In short-term exposure protocols BALB/cJ mice received a single HDM challenge (100 μg)+ isotype control, HDM challenge + HDL (200μg) and isotype control, or HDM + αSAAab and were sacrificed 24 h later (used in Fig. 2e). Contol mice received either PBS + isotype or PBS + HDL and isotype control. (e) For overexpression of SAA1 in vivo, mice were injected 20 µg of DNA complexed to polyethylenimine at day 0, exposed to PBS or HDM 48 h later and ILC2s as well as BAL cytokines measurements were performed on day 3 (used Fig. 2f). (f) WT and Saa–/– mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 μg with extracts from the parasitic worm Schistosoma mansoni (a Puerto Rican isolate). Tissues were harvested 72 h after the last allergen challenge (used in Fig. 4). (g) For FPR2 blockade (WRW4, 2 mg/kg), WT BALB/cJ mice were sensitized and challenged i.t. on day 0 and 14 with 100 μg of HDM extract. Airway measurements were performed 72 h after the last allergen challenge (used in Fig. 7a-​-f).f). (h) In short term exposure experiments, BALB/cJ mice received a single HDM challenge (100 μg) or HDM + WRW4 and were sacrificed 24 h later (used in Fig. 7g and ​andi).i). (i) Model of Alternaria alternata (Alt a )-induced airway inflammation for WT and Saa–/– mice. Mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 µg of Alt a extract. Tissues were harvested 72 h after the last allergen challenge (used in Extended Data Fig. 9f-​-jj).

Journal: Nature immunology

Article Title: Serum Amyloid A is a Soluble Pattern Recognition Receptor that Drives Type 2 Immunity

doi: 10.1038/s41590-020-0698-1

Figure Lengend Snippet: (a) Model of allergen-induced airway hyperresponsiveness (AHR) for wild-type (WT) and Saa–/– mice (both C57BL/6 background). Mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 μg of HDM extract. Airway measurements were performed 72 h after the last allergen challenge (used in Fig. 1a-​-jj and Extended Data Fig. 2 and ​and3).3). (b) For SAA1 antibody blockade, we used an established mouse model of allergen-induced AHR sensitizing WT BALB/cJ mice i.t. on day 0 and 14 with 100 µg of HDM extract + isotype control, or HDM + αSAAab. Airway measurements and tissue harvests were performed 72 h after the last allergen challenge (used in Extended Data Fig. 4). (c) In short-term exposure protocols WT and Saa–/– mice (both C57BL/6 background) received a single HDM challenge (100 μg) were sacrificed 16 h later (used in Fig. 2a-​-d).d). (d) In short-term exposure protocols BALB/cJ mice received a single HDM challenge (100 μg)+ isotype control, HDM challenge + HDL (200μg) and isotype control, or HDM + αSAAab and were sacrificed 24 h later (used in Fig. 2e). Contol mice received either PBS + isotype or PBS + HDL and isotype control. (e) For overexpression of SAA1 in vivo, mice were injected 20 µg of DNA complexed to polyethylenimine at day 0, exposed to PBS or HDM 48 h later and ILC2s as well as BAL cytokines measurements were performed on day 3 (used Fig. 2f). (f) WT and Saa–/– mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 μg with extracts from the parasitic worm Schistosoma mansoni (a Puerto Rican isolate). Tissues were harvested 72 h after the last allergen challenge (used in Fig. 4). (g) For FPR2 blockade (WRW4, 2 mg/kg), WT BALB/cJ mice were sensitized and challenged i.t. on day 0 and 14 with 100 μg of HDM extract. Airway measurements were performed 72 h after the last allergen challenge (used in Fig. 7a-​-f).f). (h) In short term exposure experiments, BALB/cJ mice received a single HDM challenge (100 μg) or HDM + WRW4 and were sacrificed 24 h later (used in Fig. 7g and ​andi).i). (i) Model of Alternaria alternata (Alt a )-induced airway inflammation for WT and Saa–/– mice. Mice were sensitized i.t. on day 0 (1 μg) and i.n. on days 7–11 with 10 µg of Alt a extract. Tissues were harvested 72 h after the last allergen challenge (used in Extended Data Fig. 9f-​-jj).

Article Snippet: HDL-depleted supernatants as well as HDL-precipitates were analyzed for their SAA1 content by immunoblotting; immunoreactivity was tested with mouse anti-human SAA1 antibody (DM1004B, Acris; MAB30196, R&D Systems) or sequence-specific antibodies as described previously 30 .

Techniques: Control, Over Expression, In Vivo, Injection

Basal (a) SAA1 and (b) FPR2 mRNA expression in nasal epithelial cells of CRS patients and matched control donors. (c) HDM (100 μg/ml)-triggered IL-33 concentration in primary nasal epithelial cells of CRS patients as compared to controls. (d) Dissociation of hexameric SAA was analyzed separating lipid-free and lipid-bound SAA by HDL pull down using a polyclonal goat antibody specific for human ApoA1 and immunoblotting with a monoclonal mouse antibody specific for human SAA1 (Acris). (e) Bar graph represents quantitative analysis of SAA monomer band intensities (LI-COR Image Studio Software). (f) SAA1 protein amounts in sera of control donors and HDM allergic individuals. Data represents means ± SEM of (a and b) n=16 control and n=27 CRS, (c) n=6 control and n=12 CRS patients per group, (e) n=5 control and n=8 CRS patients per group and (f) n=18 control and n=27 HDM allergic patients per group. Representative immunoblot of one control and one CRS patient (d). Cropped images are shown. P values were calculated with a two-tailed test using Student’s t-test with Welch’s correction. ****P ≤ 0.0001.

Journal: Nature immunology

Article Title: Serum Amyloid A is a Soluble Pattern Recognition Receptor that Drives Type 2 Immunity

doi: 10.1038/s41590-020-0698-1

Figure Lengend Snippet: Basal (a) SAA1 and (b) FPR2 mRNA expression in nasal epithelial cells of CRS patients and matched control donors. (c) HDM (100 μg/ml)-triggered IL-33 concentration in primary nasal epithelial cells of CRS patients as compared to controls. (d) Dissociation of hexameric SAA was analyzed separating lipid-free and lipid-bound SAA by HDL pull down using a polyclonal goat antibody specific for human ApoA1 and immunoblotting with a monoclonal mouse antibody specific for human SAA1 (Acris). (e) Bar graph represents quantitative analysis of SAA monomer band intensities (LI-COR Image Studio Software). (f) SAA1 protein amounts in sera of control donors and HDM allergic individuals. Data represents means ± SEM of (a and b) n=16 control and n=27 CRS, (c) n=6 control and n=12 CRS patients per group, (e) n=5 control and n=8 CRS patients per group and (f) n=18 control and n=27 HDM allergic patients per group. Representative immunoblot of one control and one CRS patient (d). Cropped images are shown. P values were calculated with a two-tailed test using Student’s t-test with Welch’s correction. ****P ≤ 0.0001.

Article Snippet: HDL-depleted supernatants as well as HDL-precipitates were analyzed for their SAA1 content by immunoblotting; immunoreactivity was tested with mouse anti-human SAA1 antibody (DM1004B, Acris; MAB30196, R&D Systems) or sequence-specific antibodies as described previously 30 .

Techniques: Expressing, Control, Concentration Assay, Western Blot, Software, Two Tailed Test

(a) AHR (*P < 0.0105), (b) total serum IgE concentrations (*P = 0.0265), (c) eosinophil infiltration into the lungs, and (d) PAS stained lung sections of isotype (iso), HDM+isotype (HDM), or HDM+αSAAab-treated (αSAA) WT BALB/c mice. Antibodies were administered at 5 μg/i.t. Frequency of (e) Lin-CD45+ST2+IL-13+ ILC2s (**P = 0.0027, ***P = 0.0002), (f) TH2 and (*P = 0.0260, ***P = 0.0001) (g) TH17 cells (*P = 0.0149, ***P = 0.0005) in the lungs of these mice. Data represents means ± SEM of pooled data from 2 independent experiments containing (a) n=6 PBS+iso, n=7 HDM+iso, n=9 HDM+αSAA animals per group; (b, c, f, g) n=9 PBS+iso, n=11 HDM+iso, n=13 HDM+αSAA animals per group or are representative of 2 independent experiments with (e) n=4 PBS+iso, n=5 HDM+iso, n=7 HDM+αSAA animals per group. P values were calculated with a two-tailed test using one-way analysis of variance (ANOVA) with Dunett’s post hoc analysis that compares HDM to iso and αSAA counterparts. ****P ≤ 0.0001

Journal: Nature immunology

Article Title: Serum Amyloid A is a Soluble Pattern Recognition Receptor that Drives Type 2 Immunity

doi: 10.1038/s41590-020-0698-1

Figure Lengend Snippet: (a) AHR (*P < 0.0105), (b) total serum IgE concentrations (*P = 0.0265), (c) eosinophil infiltration into the lungs, and (d) PAS stained lung sections of isotype (iso), HDM+isotype (HDM), or HDM+αSAAab-treated (αSAA) WT BALB/c mice. Antibodies were administered at 5 μg/i.t. Frequency of (e) Lin-CD45+ST2+IL-13+ ILC2s (**P = 0.0027, ***P = 0.0002), (f) TH2 and (*P = 0.0260, ***P = 0.0001) (g) TH17 cells (*P = 0.0149, ***P = 0.0005) in the lungs of these mice. Data represents means ± SEM of pooled data from 2 independent experiments containing (a) n=6 PBS+iso, n=7 HDM+iso, n=9 HDM+αSAA animals per group; (b, c, f, g) n=9 PBS+iso, n=11 HDM+iso, n=13 HDM+αSAA animals per group or are representative of 2 independent experiments with (e) n=4 PBS+iso, n=5 HDM+iso, n=7 HDM+αSAA animals per group. P values were calculated with a two-tailed test using one-way analysis of variance (ANOVA) with Dunett’s post hoc analysis that compares HDM to iso and αSAA counterparts. ****P ≤ 0.0001

Article Snippet: HDL-depleted supernatants as well as HDL-precipitates were analyzed for their SAA1 content by immunoblotting; immunoreactivity was tested with mouse anti-human SAA1 antibody (DM1004B, Acris; MAB30196, R&D Systems) or sequence-specific antibodies as described previously 30 .

Techniques: Neutralization, Staining, Two Tailed Test

Increase of the type 2 cytokines (a) IL-33 (*P = 0.0138), (b) IL-25, and (c) TSLP in the BAL of WT and Saa–/– mice 24h after a single dose of PBS or HDM. (d) Numbers of IL-13+ ILC2 cells in the lungs of these mice. (e) Effects of local SAA1 neutralization in WT BALB/c mice through HDL (*P = 0.0117) or local SAA antibody blockade (αSAA, *P = 0.0392) of WT BALB/c receiving isotype (iso), HDM+isotype (HDM+iso), or HDM+αSAAab-treated (HDM+αSAA). (f) Effects of SAA1 overexpression on numbers of Lin-CD45+CD25+ST2+IL-13+ ILC2s in the lungs. For overexpression, WT mice were injected retro-orbitally with a SAA1 overexpression plasmid (SAA1, grey bars) or a non-coding control vector (pcDNA, open bars). Data represent means ± SEM of pooled data from 2 independent experiments containing (a-c) n= 6 WT PBS, n=8 WT HDM, n=7 Saa–/– PBS and n=10 Saa–/– HDM animals per group; (d) n= 5 WT PBS, n=9 WT HDM, n=6 Saa–/– PBS and n=9 Saa–/– HDM animals per group; (e) n=7 PBS+iso, n=6 PBS+iso/HDL, n= 11 HDM+iso, n=13 HDM+iso/HDL, n=12 HDM+αSAA animals per group; (f) n=3 pcDNA PBS, n=9 pcDNA HDM, n=3 SAA1 PBS and n=10 SAA1 HDM animals per group. P values were calculated with a two-tailed test using one-way analysis of variance (ANOVA) with Dunett’s post hoc analysis (a-e) or two-sided Student’s t-test (f). ***P ≤ 0.001

Journal: Nature immunology

Article Title: Serum Amyloid A is a Soluble Pattern Recognition Receptor that Drives Type 2 Immunity

doi: 10.1038/s41590-020-0698-1

Figure Lengend Snippet: Increase of the type 2 cytokines (a) IL-33 (*P = 0.0138), (b) IL-25, and (c) TSLP in the BAL of WT and Saa–/– mice 24h after a single dose of PBS or HDM. (d) Numbers of IL-13+ ILC2 cells in the lungs of these mice. (e) Effects of local SAA1 neutralization in WT BALB/c mice through HDL (*P = 0.0117) or local SAA antibody blockade (αSAA, *P = 0.0392) of WT BALB/c receiving isotype (iso), HDM+isotype (HDM+iso), or HDM+αSAAab-treated (HDM+αSAA). (f) Effects of SAA1 overexpression on numbers of Lin-CD45+CD25+ST2+IL-13+ ILC2s in the lungs. For overexpression, WT mice were injected retro-orbitally with a SAA1 overexpression plasmid (SAA1, grey bars) or a non-coding control vector (pcDNA, open bars). Data represent means ± SEM of pooled data from 2 independent experiments containing (a-c) n= 6 WT PBS, n=8 WT HDM, n=7 Saa–/– PBS and n=10 Saa–/– HDM animals per group; (d) n= 5 WT PBS, n=9 WT HDM, n=6 Saa–/– PBS and n=9 Saa–/– HDM animals per group; (e) n=7 PBS+iso, n=6 PBS+iso/HDL, n= 11 HDM+iso, n=13 HDM+iso/HDL, n=12 HDM+αSAA animals per group; (f) n=3 pcDNA PBS, n=9 pcDNA HDM, n=3 SAA1 PBS and n=10 SAA1 HDM animals per group. P values were calculated with a two-tailed test using one-way analysis of variance (ANOVA) with Dunett’s post hoc analysis (a-e) or two-sided Student’s t-test (f). ***P ≤ 0.001

Article Snippet: HDL-depleted supernatants as well as HDL-precipitates were analyzed for their SAA1 content by immunoblotting; immunoreactivity was tested with mouse anti-human SAA1 antibody (DM1004B, Acris; MAB30196, R&D Systems) or sequence-specific antibodies as described previously 30 .

Techniques: Neutralization, Over Expression, Injection, Plasmid Preparation, Control, Two Tailed Test

(a) HDM, rBlo t 13, rDer p2 or rDer p 23 were separated by SDS-PAGE followed by detection with rSAA1 and a mouse monoclonal antibody specific for human SAA1. (b) Migration of SAA1 (1 mg/ml) in PBS was analyzed in the presence of increasing amounts of the mite FABP Blo t 13 (4:1 (0.25 mg/ml), 2:1 (0.5 mg/ml), 1:1 (1 mg/ml) and 1:2 (2 mg/ml) of Blo t 13) by native PAGE followed by immunoblot analysis using mouse monoclonal antibody specific for human SAA1 (R&D systems; MAB30196). (c) SAA1 (1 mg/ml) was chemically cross-linked in the presence of Blo t 13 added at a ratio of 1:2 or 1:4 (as indicated) using 0.0025%, 0.005%, and 0.01% glutaraldehyde and analysed by immunoblot using sequence-specific (amino acid 14–30) rabbit antiserum for human SAA1 (densitometric analysis shown in supplementary table 1). (d) HDM-induced IL-33 concentrations in BEAS-2B cells after siRNA-mediated silencing of SAA1 (siSAA1) or non-targeting scrambled siRNA (siNT) (**P = 0.0002, ns = 0.1911). (e) IL-33 amounts induced by individual mite or unrelated major cat (Fel d 1) and birch pollen (Bet v 1) allergens (10 μg/ml) (. (f) Blo t 13-induced IL-33 in BEAS-2B cells with siRNA-mediated silencing of SAA1 (siSAA1) or transfected with non-targeting scrambled siRNA (siNT). (g) IL-33 amounts induced by Der p 13-depleted HDM extract and HDM extract where Der p 13 was neutralized (100 μg/ml). (h) BAL IL-33 levels (*P = 0.0209, ***P = 0.006) as well as (i) Lin-CD45+ST2+IL-13+ ILC2s in the lungs of WT BALB/c mice receiving a single i.t. challenge with 100 μg Der p 13-depleted HDM extract as compared to isotype-treated control extract (*P = 0.0423). IL-33 amounts induced in the BEAS-2B cell line and SAA1 binding by the human fatty acid binding proteins (j) FABP5 (**P = 0.0207) and (k) FABP7. Cropped images are shown. Data are shown as means ± SEM and are pooled data from 2 (d, e, g, j) or 3 (h, i) independent experiments or representative of 2 (k) or 3 independent experiments (f) each containing at least n=4 biologically independent samples or n=8 PBS, n=12 HDM-isotype and n= 14 HDM-α-group 13 (h) and or n=10 PBS, n=13 HDM-isotype and n= 14 HDM-α-group 13 (i) animals per group. Immunoblots are representative of an experimental n=2 (a-c, j, k). P values were calculated with a two-tailed test using one-way analysis of variance (ANOVA) with Tukey’s multiple comparison test (d, e, f), Dunett’s (h) or Holm-Sidaks (i) post hoc analysis, and two-tailed Student’s t-test with Welch’s correction (g, j, k) ****P ≤ 0.0001.

Journal: Nature immunology

Article Title: Serum Amyloid A is a Soluble Pattern Recognition Receptor that Drives Type 2 Immunity

doi: 10.1038/s41590-020-0698-1

Figure Lengend Snippet: (a) HDM, rBlo t 13, rDer p2 or rDer p 23 were separated by SDS-PAGE followed by detection with rSAA1 and a mouse monoclonal antibody specific for human SAA1. (b) Migration of SAA1 (1 mg/ml) in PBS was analyzed in the presence of increasing amounts of the mite FABP Blo t 13 (4:1 (0.25 mg/ml), 2:1 (0.5 mg/ml), 1:1 (1 mg/ml) and 1:2 (2 mg/ml) of Blo t 13) by native PAGE followed by immunoblot analysis using mouse monoclonal antibody specific for human SAA1 (R&D systems; MAB30196). (c) SAA1 (1 mg/ml) was chemically cross-linked in the presence of Blo t 13 added at a ratio of 1:2 or 1:4 (as indicated) using 0.0025%, 0.005%, and 0.01% glutaraldehyde and analysed by immunoblot using sequence-specific (amino acid 14–30) rabbit antiserum for human SAA1 (densitometric analysis shown in supplementary table 1). (d) HDM-induced IL-33 concentrations in BEAS-2B cells after siRNA-mediated silencing of SAA1 (siSAA1) or non-targeting scrambled siRNA (siNT) (**P = 0.0002, ns = 0.1911). (e) IL-33 amounts induced by individual mite or unrelated major cat (Fel d 1) and birch pollen (Bet v 1) allergens (10 μg/ml) (. (f) Blo t 13-induced IL-33 in BEAS-2B cells with siRNA-mediated silencing of SAA1 (siSAA1) or transfected with non-targeting scrambled siRNA (siNT). (g) IL-33 amounts induced by Der p 13-depleted HDM extract and HDM extract where Der p 13 was neutralized (100 μg/ml). (h) BAL IL-33 levels (*P = 0.0209, ***P = 0.006) as well as (i) Lin-CD45+ST2+IL-13+ ILC2s in the lungs of WT BALB/c mice receiving a single i.t. challenge with 100 μg Der p 13-depleted HDM extract as compared to isotype-treated control extract (*P = 0.0423). IL-33 amounts induced in the BEAS-2B cell line and SAA1 binding by the human fatty acid binding proteins (j) FABP5 (**P = 0.0207) and (k) FABP7. Cropped images are shown. Data are shown as means ± SEM and are pooled data from 2 (d, e, g, j) or 3 (h, i) independent experiments or representative of 2 (k) or 3 independent experiments (f) each containing at least n=4 biologically independent samples or n=8 PBS, n=12 HDM-isotype and n= 14 HDM-α-group 13 (h) and or n=10 PBS, n=13 HDM-isotype and n= 14 HDM-α-group 13 (i) animals per group. Immunoblots are representative of an experimental n=2 (a-c, j, k). P values were calculated with a two-tailed test using one-way analysis of variance (ANOVA) with Tukey’s multiple comparison test (d, e, f), Dunett’s (h) or Holm-Sidaks (i) post hoc analysis, and two-tailed Student’s t-test with Welch’s correction (g, j, k) ****P ≤ 0.0001.

Article Snippet: HDL-depleted supernatants as well as HDL-precipitates were analyzed for their SAA1 content by immunoblotting; immunoreactivity was tested with mouse anti-human SAA1 antibody (DM1004B, Acris; MAB30196, R&D Systems) or sequence-specific antibodies as described previously 30 .

Techniques: SDS Page, Migration, Clear Native PAGE, Western Blot, Sequencing, Transfection, Control, Binding Assay, Two Tailed Test, Comparison

(a) Migration of SAA1 (1 mg/ml) in IMDM media was analyzed in the presence of the mite FABP rBlo t 13 (1 mg/ml) by native PAGE followed by immunoblot analysis using a sequence-specific antiserum (amino acid 89–104) raised against human SAA1. (b) Effects of the protein synthesis inhibitor cycloheximide on IL-33 concentrations in BEAS-2B cells treated for 30 min with HDM (100 μg/ml). (c) Cell viability of BEAS-2B cells after HDM exposure over time as measured by continuous reduction of a cell viability substrate by viable cells (**P = 0.0041). SAA1 concentrations in cells after (d) siRNA-mediated silencing of SAA1 (siSAA1) or non-targeting scrambled siRNA (siNT). Effects of SAA1 on HDM-induced IL-6 (**P = 0.0086, ***P = 0.0007) and IL-8 release in cells with siRNA mediated knockdown of SAA1 (siSAA1) (e and f). (g) HDM-induced IL-6 amounts after Der p 13-depletion and/or neutralization. Data are shown as means ± SEM and are representative of 2 (b, e) or 3 (c) independent experiments or pooled data from 2 independent experiments (d, f) each containing at least n=4 biologically independent samples. Representative analysis of SAA1 migration patterns in the presence of Blo t 13 using sequence-specific rabbit antiserum raised against human SAA1 (aa 89–104) (a). IL-6 amounts were analysed for one representative experiments with n=5 biologically independent samples (g). Cropped images are shown. P values were calculated with a two-tailed test using one-way analysis of variance (ANOVA) followed by Dunett’s post (b) or Tukey’s hoc analysis (e,f), two-way ANOVA followed by Dunnet correction (c) or Student’s t-test (d, g). ***P ≤ 0.001; ****P ≤ 0.0001.

Journal: Nature immunology

Article Title: Serum Amyloid A is a Soluble Pattern Recognition Receptor that Drives Type 2 Immunity

doi: 10.1038/s41590-020-0698-1

Figure Lengend Snippet: (a) Migration of SAA1 (1 mg/ml) in IMDM media was analyzed in the presence of the mite FABP rBlo t 13 (1 mg/ml) by native PAGE followed by immunoblot analysis using a sequence-specific antiserum (amino acid 89–104) raised against human SAA1. (b) Effects of the protein synthesis inhibitor cycloheximide on IL-33 concentrations in BEAS-2B cells treated for 30 min with HDM (100 μg/ml). (c) Cell viability of BEAS-2B cells after HDM exposure over time as measured by continuous reduction of a cell viability substrate by viable cells (**P = 0.0041). SAA1 concentrations in cells after (d) siRNA-mediated silencing of SAA1 (siSAA1) or non-targeting scrambled siRNA (siNT). Effects of SAA1 on HDM-induced IL-6 (**P = 0.0086, ***P = 0.0007) and IL-8 release in cells with siRNA mediated knockdown of SAA1 (siSAA1) (e and f). (g) HDM-induced IL-6 amounts after Der p 13-depletion and/or neutralization. Data are shown as means ± SEM and are representative of 2 (b, e) or 3 (c) independent experiments or pooled data from 2 independent experiments (d, f) each containing at least n=4 biologically independent samples. Representative analysis of SAA1 migration patterns in the presence of Blo t 13 using sequence-specific rabbit antiserum raised against human SAA1 (aa 89–104) (a). IL-6 amounts were analysed for one representative experiments with n=5 biologically independent samples (g). Cropped images are shown. P values were calculated with a two-tailed test using one-way analysis of variance (ANOVA) followed by Dunett’s post (b) or Tukey’s hoc analysis (e,f), two-way ANOVA followed by Dunnet correction (c) or Student’s t-test (d, g). ***P ≤ 0.001; ****P ≤ 0.0001.

Article Snippet: HDL-depleted supernatants as well as HDL-precipitates were analyzed for their SAA1 content by immunoblotting; immunoreactivity was tested with mouse anti-human SAA1 antibody (DM1004B, Acris; MAB30196, R&D Systems) or sequence-specific antibodies as described previously 30 .

Techniques: Derivative Assay, Migration, Clear Native PAGE, Western Blot, Sequencing, Knockdown, Neutralization, Two Tailed Test

(a) Sequence of SAA1. Indicated are: secondary structure α-helices (α 1–4) and loops (connecting lines between α-helices), C-terminal tail (CTL). Amino acids located within the hydrophobic core of the SAA1 hexamer are shaded in green. The epitopes recognized by affinity-purified IgGs raised against residues 27–44, 40–63, 68–84, and 89–104 are indicated by yellow lines (used in panel e). The mutation site at the hydrophobic core is indicated by an orange dot (used in panel f). C-terminal deletion of amino acids (Δ1–11) is shaded in blue. (b) BEAS-2B supernatants were cleared of lipid-bound SAA1 pulling down HDL-bound SAA1 using a polyclonal goat anti-ApoA1 antibody. Cleared supernatants were immunoblotted with a monoclonal mouse anti-human SAA1 antibody (Acris). (c) IL-33 secretion induced by rSAA1 alone (open bars) or in complex with a mouse monoclonal antibody specific for human SAA1 (αSAA ab, closed bars) in the BEAS-2B cell line. (d) Supernatants of cells left either untreated (media) or stimulated with recombinant SAA1 (rSAA) and immunoblotted with a monoclonal antibody specific for human SAA (R&D Systems; MAB30196). (e) IL-33 release from BEAS-2B cells after incubation with sequence-specific rabbit antisera specific for human SAA1 (**P = 0.0029). (f) HDM-triggered IL-33 release in BEAS-2B cells transfected with empty plasmid control (EV), wildtype SAA1 (WT) overexpression plasmid or a SAA1 plasmid with a Trp to Ala mutation at position 53 of the amino acid sequence (W53A) to mutate the hydrophobic core of SAA1 (WT to EV **P = 0.0032; WT to W53A **P = 0.0085). (g) HDM-induced IL-33 amounts in epithelial cells grown in media supplemented with charcoal-stripped FBS (**P = 0.0027). Cropped images are shown. Data shown as means ± SEM represent are representative of 2 independent experiments (c, d, f, g) or 3 (e) each containing at least n=4 biologically independent samples. Blots are representative of 2 (d) and 4 (b) independent experiments. P values were calculated with a two-tailed test using one-way analysis of variance (ANOVA) with Tukey’s multiple comparison test (e, g) or Dunett’s post hoc analysis (f). ****P ≤ 0.0001

Journal: Nature immunology

Article Title: Serum Amyloid A is a Soluble Pattern Recognition Receptor that Drives Type 2 Immunity

doi: 10.1038/s41590-020-0698-1

Figure Lengend Snippet: (a) Sequence of SAA1. Indicated are: secondary structure α-helices (α 1–4) and loops (connecting lines between α-helices), C-terminal tail (CTL). Amino acids located within the hydrophobic core of the SAA1 hexamer are shaded in green. The epitopes recognized by affinity-purified IgGs raised against residues 27–44, 40–63, 68–84, and 89–104 are indicated by yellow lines (used in panel e). The mutation site at the hydrophobic core is indicated by an orange dot (used in panel f). C-terminal deletion of amino acids (Δ1–11) is shaded in blue. (b) BEAS-2B supernatants were cleared of lipid-bound SAA1 pulling down HDL-bound SAA1 using a polyclonal goat anti-ApoA1 antibody. Cleared supernatants were immunoblotted with a monoclonal mouse anti-human SAA1 antibody (Acris). (c) IL-33 secretion induced by rSAA1 alone (open bars) or in complex with a mouse monoclonal antibody specific for human SAA1 (αSAA ab, closed bars) in the BEAS-2B cell line. (d) Supernatants of cells left either untreated (media) or stimulated with recombinant SAA1 (rSAA) and immunoblotted with a monoclonal antibody specific for human SAA (R&D Systems; MAB30196). (e) IL-33 release from BEAS-2B cells after incubation with sequence-specific rabbit antisera specific for human SAA1 (**P = 0.0029). (f) HDM-triggered IL-33 release in BEAS-2B cells transfected with empty plasmid control (EV), wildtype SAA1 (WT) overexpression plasmid or a SAA1 plasmid with a Trp to Ala mutation at position 53 of the amino acid sequence (W53A) to mutate the hydrophobic core of SAA1 (WT to EV **P = 0.0032; WT to W53A **P = 0.0085). (g) HDM-induced IL-33 amounts in epithelial cells grown in media supplemented with charcoal-stripped FBS (**P = 0.0027). Cropped images are shown. Data shown as means ± SEM represent are representative of 2 independent experiments (c, d, f, g) or 3 (e) each containing at least n=4 biologically independent samples. Blots are representative of 2 (d) and 4 (b) independent experiments. P values were calculated with a two-tailed test using one-way analysis of variance (ANOVA) with Tukey’s multiple comparison test (e, g) or Dunett’s post hoc analysis (f). ****P ≤ 0.0001

Article Snippet: HDL-depleted supernatants as well as HDL-precipitates were analyzed for their SAA1 content by immunoblotting; immunoreactivity was tested with mouse anti-human SAA1 antibody (DM1004B, Acris; MAB30196, R&D Systems) or sequence-specific antibodies as described previously 30 .

Techniques: Sequencing, Affinity Purification, Mutagenesis, Recombinant, Incubation, Transfection, Plasmid Preparation, Control, Over Expression, Two Tailed Test, Comparison

(a) mRNA and (b) protein amounts of SAA in response to HDM (100 μg/ml). (c) SAA1 hexamer after rBlo t 13 stimulation was analyzed as described in Fig. 3b. (d) Bar graph represents quantitative analysis of SAA hexamer using LI-COR Image Studio Software. (e) Concentration-dependent IL-33 release from BEAS-2B cells induced by rBlo t 13. Data are shown as means ± SEM and are pooled data from 2 independent experiments (b, e) each containing n=4–5 replicate wells or representative of 2–3 independent experiments (d). SAA1 mRNA expression, normalized to the average of housekeeping genes, is presented as mean value ± SEM (n = 5). Immunoblot is representative of an experimental n=2. Cropped images are shown. P values were calculated with a two-tailed test using Student’s t-test (a), two-way analysis of variance (ANOVA) followed by Dunnet’s correction (b) or one-way ANOVA with Dunett’s post hoc analysis (d, e). ****P ≤ 0.0001.

Journal: Nature immunology

Article Title: Serum Amyloid A is a Soluble Pattern Recognition Receptor that Drives Type 2 Immunity

doi: 10.1038/s41590-020-0698-1

Figure Lengend Snippet: (a) mRNA and (b) protein amounts of SAA in response to HDM (100 μg/ml). (c) SAA1 hexamer after rBlo t 13 stimulation was analyzed as described in Fig. 3b. (d) Bar graph represents quantitative analysis of SAA hexamer using LI-COR Image Studio Software. (e) Concentration-dependent IL-33 release from BEAS-2B cells induced by rBlo t 13. Data are shown as means ± SEM and are pooled data from 2 independent experiments (b, e) each containing n=4–5 replicate wells or representative of 2–3 independent experiments (d). SAA1 mRNA expression, normalized to the average of housekeeping genes, is presented as mean value ± SEM (n = 5). Immunoblot is representative of an experimental n=2. Cropped images are shown. P values were calculated with a two-tailed test using Student’s t-test (a), two-way analysis of variance (ANOVA) followed by Dunnet’s correction (b) or one-way ANOVA with Dunett’s post hoc analysis (d, e). ****P ≤ 0.0001.

Article Snippet: HDL-depleted supernatants as well as HDL-precipitates were analyzed for their SAA1 content by immunoblotting; immunoreactivity was tested with mouse anti-human SAA1 antibody (DM1004B, Acris; MAB30196, R&D Systems) or sequence-specific antibodies as described previously 30 .

Techniques: Software, Concentration Assay, Expressing, Western Blot, Two Tailed Test

(a) mRNA expression of the FPR family members FPR1, FPR2 and FPR3 at baseline (open bars) or after 2 h of HDM stimulation (filled bars). (b) HDM-triggered IL-33 amounts in BEAS-2B cells overexpressing human FPR1 (**P = 0.0068, ***P = 0.0003). (c) IL-33 secretion in BEAS-2B cells overexpressing human FPR2 or cells transfected with an empty vector (EV; pcDNA3.1) (**P = 0.0068, ***P = 0.0003). HDM-induced IL-6 and IL-8 amounts in BEAS-2B cells overexpressing FPR2 (d and e) or blocking the FPR2 receptor (f (**P = 0.0043) and g (**P = 0.0021)) using WRW4. Data presented as means ± SEM and is representative of 2 independent experiments each containing at least n= 4 biologically independent samples (b, d, f) or pooled data from 2 independent experiments (c, e, g). mRNA expression, normalized to the average of housekeeping genes, is presented as mean values ± SEM (n = 5 biologically independent samples) performed in duplicates (a). P values were calculated with a two-tailed test using Student’s t-test (a) or one-way analysis of variance (ANOVA) with Tukeys multiple comparison test (b-e) or Dunett’s post hoc analysis (f, g). ****P ≤ 0.0001

Journal: Nature immunology

Article Title: Serum Amyloid A is a Soluble Pattern Recognition Receptor that Drives Type 2 Immunity

doi: 10.1038/s41590-020-0698-1

Figure Lengend Snippet: (a) mRNA expression of the FPR family members FPR1, FPR2 and FPR3 at baseline (open bars) or after 2 h of HDM stimulation (filled bars). (b) HDM-triggered IL-33 amounts in BEAS-2B cells overexpressing human FPR1 (**P = 0.0068, ***P = 0.0003). (c) IL-33 secretion in BEAS-2B cells overexpressing human FPR2 or cells transfected with an empty vector (EV; pcDNA3.1) (**P = 0.0068, ***P = 0.0003). HDM-induced IL-6 and IL-8 amounts in BEAS-2B cells overexpressing FPR2 (d and e) or blocking the FPR2 receptor (f (**P = 0.0043) and g (**P = 0.0021)) using WRW4. Data presented as means ± SEM and is representative of 2 independent experiments each containing at least n= 4 biologically independent samples (b, d, f) or pooled data from 2 independent experiments (c, e, g). mRNA expression, normalized to the average of housekeeping genes, is presented as mean values ± SEM (n = 5 biologically independent samples) performed in duplicates (a). P values were calculated with a two-tailed test using Student’s t-test (a) or one-way analysis of variance (ANOVA) with Tukeys multiple comparison test (b-e) or Dunett’s post hoc analysis (f, g). ****P ≤ 0.0001

Article Snippet: HDL-depleted supernatants as well as HDL-precipitates were analyzed for their SAA1 content by immunoblotting; immunoreactivity was tested with mouse anti-human SAA1 antibody (DM1004B, Acris; MAB30196, R&D Systems) or sequence-specific antibodies as described previously 30 .

Techniques: Expressing, Transfection, Plasmid Preparation, Blocking Assay, Two Tailed Test, Comparison

IL-33 concentrations in BEAS-2B cells blocking the SAA-binding receptors (a) FPR2 (WRW4, 12 μM; ***P = 0.0006) and (b) TLR4 (LPS from Rhodobacter sphaeroides; LPS-RS, 10 μg/ml; ). (c) Effects of FPR2 or TLR4 blockade in cells transfected with empty plasmid control (EV) or SAA1 overexpression plasmid. **P = 0.0047 § indicates p = 0.056; # indicates p = 0.22 between EV and SAA plasmid. Blockade of (d) CD36 (anti-human CD36 blocking antibody, aCD36; 10 μg/ml; ***P = 0.0005), (e) the P2 receptor antagonist suramin (100 μM; *P = 0.028), and (f) TLR2 (anti-human TLR2 IgA, aTLR2, 10 μg/ml; **P = 0.0055). (g) HDM-triggered IL-33 release in BEAS-2B cells transfected with EV, WT SAA1 overexpression plasmid or a SAA1 plasmid with a deletion of the C-terminal amino acids 1–11 (Δ1–11; *P = 0.01, **P = 0.0078 ). Data are representative of 2–3 independent experiments (b, d-f) or pooled data from 2 independent experiments (a, c, d, g) each containing at least n=4 biologically independent samples and depicted as means ± SEM. P values were calculated with a two-tailed test using one-way analysis of variance (ANOVA) with Dunett’s (a, b, d-g) or Tukey’s (c) post hoc analysis.****P ≤ 0.0001.

Journal: Nature immunology

Article Title: Serum Amyloid A is a Soluble Pattern Recognition Receptor that Drives Type 2 Immunity

doi: 10.1038/s41590-020-0698-1

Figure Lengend Snippet: IL-33 concentrations in BEAS-2B cells blocking the SAA-binding receptors (a) FPR2 (WRW4, 12 μM; ***P = 0.0006) and (b) TLR4 (LPS from Rhodobacter sphaeroides; LPS-RS, 10 μg/ml; ). (c) Effects of FPR2 or TLR4 blockade in cells transfected with empty plasmid control (EV) or SAA1 overexpression plasmid. **P = 0.0047 § indicates p = 0.056; # indicates p = 0.22 between EV and SAA plasmid. Blockade of (d) CD36 (anti-human CD36 blocking antibody, aCD36; 10 μg/ml; ***P = 0.0005), (e) the P2 receptor antagonist suramin (100 μM; *P = 0.028), and (f) TLR2 (anti-human TLR2 IgA, aTLR2, 10 μg/ml; **P = 0.0055). (g) HDM-triggered IL-33 release in BEAS-2B cells transfected with EV, WT SAA1 overexpression plasmid or a SAA1 plasmid with a deletion of the C-terminal amino acids 1–11 (Δ1–11; *P = 0.01, **P = 0.0078 ). Data are representative of 2–3 independent experiments (b, d-f) or pooled data from 2 independent experiments (a, c, d, g) each containing at least n=4 biologically independent samples and depicted as means ± SEM. P values were calculated with a two-tailed test using one-way analysis of variance (ANOVA) with Dunett’s (a, b, d-g) or Tukey’s (c) post hoc analysis.****P ≤ 0.0001.

Article Snippet: HDL-depleted supernatants as well as HDL-precipitates were analyzed for their SAA1 content by immunoblotting; immunoreactivity was tested with mouse anti-human SAA1 antibody (DM1004B, Acris; MAB30196, R&D Systems) or sequence-specific antibodies as described previously 30 .

Techniques: Blocking Assay, Binding Assay, Transfection, Plasmid Preparation, Control, Over Expression, Two Tailed Test

(a) Immunoblot of SAA1 after Alternaria alternata (Alt a) stimulation of BEAS-2B cells performed as described in Fig. 3b. Alt a-induced IL-33 and IL-8 (**P = 0.0044) secretion in BEAS-2B cells after siRNA-mediated silencing of SAA1 (siSAA1) (b and c) or WRW4-mediated FPR2 blockade (d and e). (f) Total serum IgE concentrations, (g) eosinophil counts and frequency of (h) CD3+CD4+, (i) TH2 and (j) TH17 cells in the lungs of PBS or Alt a-treated WT and Saa–/– mice. Immunoblots are representative of an experimental n=2 (a). Data are presented as means ± SEM and represent pooled data from 2 independent experiments (b, d, e, f, g, i, j) or show one representative experiment (c) each containing at least n= 4 biologically independent samples or n=8 WT PBS, n=13 WT Alt a, n=8 Saa–/– PBS and n=11 Saa–/– Alt a animals per group (f, g, i, j) or n=5 WT PBS, n=9 WT Alt a, n=5 Saa–/– PBS and n=7 Saa–/– Alt a animals per group (h). Cropped images are shown. P values were calculated with a two-tailed test using one-way analysis of variance (ANOVA) with Tukeys multiple comparison test (a, b) or Dunett’s post hoc analysis (d-j). ****P ≤ 0.0001 siNT = non-targeting siRNA; siSAA1 = SAA1-targeting siRNA. ns=not significant.

Journal: Nature immunology

Article Title: Serum Amyloid A is a Soluble Pattern Recognition Receptor that Drives Type 2 Immunity

doi: 10.1038/s41590-020-0698-1

Figure Lengend Snippet: (a) Immunoblot of SAA1 after Alternaria alternata (Alt a) stimulation of BEAS-2B cells performed as described in Fig. 3b. Alt a-induced IL-33 and IL-8 (**P = 0.0044) secretion in BEAS-2B cells after siRNA-mediated silencing of SAA1 (siSAA1) (b and c) or WRW4-mediated FPR2 blockade (d and e). (f) Total serum IgE concentrations, (g) eosinophil counts and frequency of (h) CD3+CD4+, (i) TH2 and (j) TH17 cells in the lungs of PBS or Alt a-treated WT and Saa–/– mice. Immunoblots are representative of an experimental n=2 (a). Data are presented as means ± SEM and represent pooled data from 2 independent experiments (b, d, e, f, g, i, j) or show one representative experiment (c) each containing at least n= 4 biologically independent samples or n=8 WT PBS, n=13 WT Alt a, n=8 Saa–/– PBS and n=11 Saa–/– Alt a animals per group (f, g, i, j) or n=5 WT PBS, n=9 WT Alt a, n=5 Saa–/– PBS and n=7 Saa–/– Alt a animals per group (h). Cropped images are shown. P values were calculated with a two-tailed test using one-way analysis of variance (ANOVA) with Tukeys multiple comparison test (a, b) or Dunett’s post hoc analysis (d-j). ****P ≤ 0.0001 siNT = non-targeting siRNA; siSAA1 = SAA1-targeting siRNA. ns=not significant.

Article Snippet: HDL-depleted supernatants as well as HDL-precipitates were analyzed for their SAA1 content by immunoblotting; immunoreactivity was tested with mouse anti-human SAA1 antibody (DM1004B, Acris; MAB30196, R&D Systems) or sequence-specific antibodies as described previously 30 .

Techniques: Western Blot, Two Tailed Test, Comparison

Basal (a) SAA1 and (b) FPR2 expression in bronchial epithelial from asthmatic patients and matched controls. Data represents means ± SEM of (a and b) n= 6 control and n= 6 asthmatic patients per group. x.

Journal: Nature immunology

Article Title: Serum Amyloid A is a Soluble Pattern Recognition Receptor that Drives Type 2 Immunity

doi: 10.1038/s41590-020-0698-1

Figure Lengend Snippet: Basal (a) SAA1 and (b) FPR2 expression in bronchial epithelial from asthmatic patients and matched controls. Data represents means ± SEM of (a and b) n= 6 control and n= 6 asthmatic patients per group. x.

Article Snippet: HDL-depleted supernatants as well as HDL-precipitates were analyzed for their SAA1 content by immunoblotting; immunoreactivity was tested with mouse anti-human SAA1 antibody (DM1004B, Acris; MAB30196, R&D Systems) or sequence-specific antibodies as described previously 30 .

Techniques: Expressing, Control

The schematic diagram of the experiment design. CYP2D6 cytochrome P450 2D6, AIH autoimmune hepatitis, DEPs differentially expressed proteins, iTRAQ isobaric tags for relative and absolute quantitation, SAA1 serum amyloid A 1, GO gene ontology

Journal: Journal of Translational Medicine

Article Title: Plasma proteomic analysis of autoimmune hepatitis in an improved AIH mouse model

doi: 10.1186/s12967-019-02180-3

Figure Lengend Snippet: The schematic diagram of the experiment design. CYP2D6 cytochrome P450 2D6, AIH autoimmune hepatitis, DEPs differentially expressed proteins, iTRAQ isobaric tags for relative and absolute quantitation, SAA1 serum amyloid A 1, GO gene ontology

Article Snippet: Primary antibody against SAA1 (1:100; 16721-1-AP, Proteintech, Rosemont, IL USA) was incubated overnight at 4 °C.

Techniques: Multiplex sample analysis, Quantitation Assay

Vital DEPs validation in patients with AIH. a Representative H&E staining of liver tissue section from a patient with AIH. b Plasma from patients with AIH, hepatitis B patients, hepatitis C patients, and healthy people were collected and the SAA1 expression levels in the plasma were detected by ELISA (n = 30, ***P < 0.001 vs. Control). c Immunohistochemical staining of human liver tissues for SAA1 detection are shown (n = 3, ×200 magnification). d Box plot of plasma SAA1 level in patients with AIH from different inflammation grades (n = 7 for G0–G1, n = 14 for G2, n = 8 for G3–G4, # Kruskal–Wallis nonparametric test *Mann–Whitney test). The line through the middle of each box represents the median. The length of the box represents the level range. e Box plot of plasma SAA1 level in patients with AIH from different fibrosis stages (n = 14 for S0–S1, n = 8 for S2, n = 7 for S3–S4, # Kruskal–Wallis nonparametric test *Mann–Whitney test)

Journal: Journal of Translational Medicine

Article Title: Plasma proteomic analysis of autoimmune hepatitis in an improved AIH mouse model

doi: 10.1186/s12967-019-02180-3

Figure Lengend Snippet: Vital DEPs validation in patients with AIH. a Representative H&E staining of liver tissue section from a patient with AIH. b Plasma from patients with AIH, hepatitis B patients, hepatitis C patients, and healthy people were collected and the SAA1 expression levels in the plasma were detected by ELISA (n = 30, ***P < 0.001 vs. Control). c Immunohistochemical staining of human liver tissues for SAA1 detection are shown (n = 3, ×200 magnification). d Box plot of plasma SAA1 level in patients with AIH from different inflammation grades (n = 7 for G0–G1, n = 14 for G2, n = 8 for G3–G4, # Kruskal–Wallis nonparametric test *Mann–Whitney test). The line through the middle of each box represents the median. The length of the box represents the level range. e Box plot of plasma SAA1 level in patients with AIH from different fibrosis stages (n = 14 for S0–S1, n = 8 for S2, n = 7 for S3–S4, # Kruskal–Wallis nonparametric test *Mann–Whitney test)

Article Snippet: Primary antibody against SAA1 (1:100; 16721-1-AP, Proteintech, Rosemont, IL USA) was incubated overnight at 4 °C.

Techniques: Biomarker Discovery, Staining, Clinical Proteomics, Expressing, Enzyme-linked Immunosorbent Assay, Control, Immunohistochemical staining, MANN-WHITNEY