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: Primers used for qRT-PCR

Article Snippet: TYRO3, AXL, and MERTK levels were evaluated by Western blot as previously described ( 8 ) using the anti-human primary antibodies against TYRO3 (#MAB859, R&D Systems), AXL (#AF154, R&D Systems), and MERTK (#AF891, R&D Systems).

Techniques:

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: TYRO3, AXL, and MERTK levels were evaluated by Western blot as previously described ( 8 ) using the anti-human primary antibodies against TYRO3 (#MAB859, R&D Systems), AXL (#AF154, R&D Systems), and MERTK (#AF891, R&D Systems).

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: TYRO3, AXL, and MERTK levels were evaluated by Western blot as previously described ( 8 ) using the anti-human primary antibodies against TYRO3 (#MAB859, R&D Systems), AXL (#AF154, R&D Systems), and MERTK (#AF891, R&D Systems).

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