human pro il 1beta  (Sino Biological)

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Viral Infection sensitizes Human Fetal Membranes to Bacterial LPS by MERTK Inhibition and Inflammasome Activation 1

doi: 10.4049/jimmunol.1700870

Figure Lengend Snippet: Human FM explants were treated with (A) No treatment (NT), LPS (100ng/ml), MHV-68 (1.5×104/ml PFU) or both MHV-68 and LPS (n=7); (B) NT, LPS (100ng/ml), HSV-2 (6.4×102/ml PFU) or both HSV-2 and LPS (n=3); or (C) NT, LPS (1ng/ml), Poly(I:C) (20µg/ml) or both Poly(I:C) and LPS (n=6). (A–C) Supernatants were measured for IL-1β by ELISA. (D) Pregnant wildtype mice were injected on E8.5 with either PBS or MHV-68 (1×106 PFU), and on E15.5 with either PBS or LPS (20µg/kg). After 6hrs mice were sacrificed. The FMs were harvested and homogenized for RNA and IL-1B mRNA levels measured by qRT-PCR. *p<0.05 relative to the NT or PBS control unless otherwise indicated. Data are expressed as mean±SEM.

Article Snippet: IL-1β levels were evaluated using the anti-human primary antibodies against pro-IL-1β (# 12703, Cell Signaling) and the active form (# 2022, Cell Signaling). β-Actin was used as a loading control (Sigma).

Techniques: Enzyme-linked Immunosorbent Assay, Injection, Quantitative RT-PCR, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Viral Infection sensitizes Human Fetal Membranes to Bacterial LPS by MERTK Inhibition and Inflammasome Activation 1

doi: 10.4049/jimmunol.1700870

Figure Lengend Snippet: (A) Human FM explants were treated with (A) No treatment (NT), LPS (100ng/ml), MHV-68 (1.5×104/ml PFU) or both MHV-68 and LPS (n=7). Lysates were evaluated by Western blot for expression of pro-IL-1β (31kDa) and the 17kDa active form. Blots are from one representative experiment. Bar charts show pro- and active-IL-1β expression as determined by densitometry and normalized to β-actin (n=4–5). (B & C) Human FM explants were treated with NT, LPS, MHV-68 or both MHV-68 and LPS in the presence of either media or (B) a caspase-1 inhibitor (1µM) (n=6); or (C) the NLRP3 inhibitor, MNS (10µM) (n=5). Supernatants were measured for IL-1β by ELISA. *p<0.05 relative to the NT control unless otherwise indicated. Data are expressed as mean±SEM.

Article Snippet: IL-1β levels were evaluated using the anti-human primary antibodies against pro-IL-1β (# 12703, Cell Signaling) and the active form (# 2022, Cell Signaling). β-Actin was used as a loading control (Sigma).

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Viral Infection sensitizes Human Fetal Membranes to Bacterial LPS by MERTK Inhibition and Inflammasome Activation 1

doi: 10.4049/jimmunol.1700870

Figure Lengend Snippet: Summary of the effect of viral infection or Poly(I:C) on human FM responses to LPS.

Article Snippet: IL-1β levels were evaluated using the anti-human primary antibodies against pro-IL-1β (# 12703, Cell Signaling) and the active form (# 2022, Cell Signaling). β-Actin was used as a loading control (Sigma).

Techniques: Infection

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Viral Infection sensitizes Human Fetal Membranes to Bacterial LPS by MERTK Inhibition and Inflammasome Activation 1

doi: 10.4049/jimmunol.1700870

Figure Lengend Snippet: Human FM explants were treated with no treatment (NT), LPS (100ng/ml), MHV-68 (1.5×104/ml PFU) or both MHV-68 and LPS (n=4); Supernatants were measured by multiplex analysis. *p<0.05 relative to the NT control unless otherwise indicated. Data are expressed as mean±SEM.

Article Snippet: IL-1β levels were evaluated using the anti-human primary antibodies against pro-IL-1β (# 12703, Cell Signaling) and the active form (# 2022, Cell Signaling). β-Actin was used as a loading control (Sigma).

Techniques: Multiplex Assay, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Viral Infection sensitizes Human Fetal Membranes to Bacterial LPS by MERTK Inhibition and Inflammasome Activation 1

doi: 10.4049/jimmunol.1700870

Figure Lengend Snippet: (A) Untreated FM explants (n=3) were homogenized and analyzed for expression of TYRO3, AXL, MERTK, GAS6 and PROS1 mRNA by qRT-PCR. (B–C) Human FM explants were treated with no treatment (NT), LPS (100ng/ml), MHV-68 (1.5×104/ml PFU) or both MHV-68 and LPS in the presence of media or rGas6 (50ng/ml) (n=5). Tissues were homogenized for protein and Western blot performed for (B) AXL (~140kDa) and (C) MERTK (~180kDa). Blots are from one representative experiment. Bar charts show AXL and MERTK expression as determined by densitometry and normalized to b-actin. (D) FM explants were treated with NT, LPS (1ng/ml), Poly(I:C) (20µg/ml) or both Poly(I:C) and LPS (n=4). Tissues were homogenized for protein and Western blot performed for AXL and MERTK. Blots are from one representative experiment. Bar charts show AXL and MERTK expression as determined by densitometry and normalized to β-actin. (E–F) Human FM explants were treated with no treatment (NT), LPS, MHV-68 or both MHV-68 and LPS in either the presence of media or rGAS6. Tissues were homogenized for protein and ELISA performed for (E) sMERTK (n=7); (F) GAS6 (n=5), and (G) PROS1 (n=8). *p<0.05 relative to the NT control unless otherwise indicated. Data are expressed as mean±SEM.

Article Snippet: IL-1β levels were evaluated using the anti-human primary antibodies against pro-IL-1β (# 12703, Cell Signaling) and the active form (# 2022, Cell Signaling). β-Actin was used as a loading control (Sigma).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Viral Infection sensitizes Human Fetal Membranes to Bacterial LPS by MERTK Inhibition and Inflammasome Activation 1

doi: 10.4049/jimmunol.1700870

Figure Lengend Snippet: (A) Human FM explants were treated with no treatment (NT), LPS (100ng/ml), MHV-68 (1.5×104/ml PFU) or both MHV-68 and LPS in the presence of media or rGas6 (50ng/ml). (i) IL-1β secretion in supernatants was measured by ELISA (n=6). Lysates were evaluated by Western blot for expression of (ii) pro-IL-1β and (iii) active IL-1β. Bar charts show pro- and active-IL-1β expression as determined by densitometry and normalized to β-actin (n=4–5) *p<0.05 relative to the NT control unless otherwise indicated. (B) Human FM explants were treated with or without LPS (1ng/ml) in the presence of blocking anti-TAM Abs or isotype control Abs. IL-1β secretion was measured and levels expressed as LPS-induced fold change (FC) relative to the NT control (n=4; *p<0.05 relative to the isotype control). (C–D) Pregnant wildtype or AXL−/−MERTK−/− mice were injected on E15.5 with either PBS or LPS (20µg/kg). After 6hrs mice were sacrificed. The FMs were harvested and homogenized for RNA. (D) TYRO3, AXL, MERTK, GAS6 and PROS1 mRNA expression levels in FMs from PBS treated wildtype mice (n=3) were measured by qRT-PCR. (E) IL-1B mRNA expression levels in FMs from PBS and LPS treated wildtype or AXL−/−MERTK−/− mice were measured by qRT-PCR. *p<0.05 relative to the control groups unless otherwise indicated. Data are expressed as mean±SEM.

Article Snippet: IL-1β levels were evaluated using the anti-human primary antibodies against pro-IL-1β (# 12703, Cell Signaling) and the active form (# 2022, Cell Signaling). β-Actin was used as a loading control (Sigma).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Control, Blocking Assay, Injection, Quantitative RT-PCR