Journal: Open Access Rheumatology : Research and Reviews

Article Title: A suppressive effect of prostaglandin E 2 on the expression of SERPINE1 /plasminogen activator inhibitor-1 in human articular chondrocytes: An in vitro pilot study

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Figure Lengend Snippet: Suppressive effect of PGE 2 on PAI-1 expression. A ) Western blot. Left: Chondrocytes were stimulated with PGE 2 (0.1, 1, or 10 nM) for 24 h, and the expression of PAI-1 was analyzed. NS: not stimulated. Representative results are shown. Bottom: Calculation of the relative intensity of the bands among four OA samples tested. B ) ELISA. Levels of PAI-1 in culture supernatants of chondrocytes after PGE 2 stimulation (0.1–10 nM) were measured. Notes: OA chondrocytes, N = 3, duplicate. Abbreviations: ELISA, enzyme-linked immunosorbent assay; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; PAI-1, plasminogen activator inhibitor; PGE 2 , prostaglandin E 2 .

Article Snippet: The level of PAI-1 in the culture supernatant was measured by using an ELISA kit (AssayMax Human Plasminogen activator inhibitor-1 ELISA Kit™, AssayPro LLC, St. Charles, MO, USA).

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

Journal: Open Access Rheumatology : Research and Reviews

Article Title: A suppressive effect of prostaglandin E 2 on the expression of SERPINE1 /plasminogen activator inhibitor-1 in human articular chondrocytes: An in vitro pilot study

doi:

Figure Lengend Snippet: The PAI-1 suppression by PGE 2 is delivered through EP4 receptor. The summary of PAI-1 ELISA with receptor antagonists are shown. Chondrocytes were stimulated with PGE 2 (10 nM) with or without AH6809 (10 ng/ml) or GW627368X (5 μM), and the levels of PAI-1 in culture supernatants were measured. Notes: OA chondrocytes, N = 6. Abbreviations: ELISA, enzyme-linked immunosorbent assay; PAI-1, plasminogen activator inhibitor; PGE 2 , prostaglandin E 2 .

Article Snippet: The level of PAI-1 in the culture supernatant was measured by using an ELISA kit (AssayMax Human Plasminogen activator inhibitor-1 ELISA Kit™, AssayPro LLC, St. Charles, MO, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Open Access Rheumatology : Research and Reviews

Article Title: A suppressive effect of prostaglandin E 2 on the expression of SERPINE1 /plasminogen activator inhibitor-1 in human articular chondrocytes: An in vitro pilot study

doi:

Figure Lengend Snippet: Inhibition of p38 and ERK MAPK did not abrogate the effect of PGE 2 on PAI-1 downregulation. The summary of PAI-1 ELISA with receptor antagonists are shown. Chondrocytes were stimulated with PGE 2 (10 nM) with or without SB203580 or PD98059, and the levels of PAI-1 in culture supernatants were measured. Notes: OA chondrocytes, N = 3. *p <0.05; **p <0.02. Abbreviations: MAPK, mitogen-activated protein kinases, PAI-1, plasminogen activator inhibitor; PGE 2 , prostaglandin E 2 .

Article Snippet: The level of PAI-1 in the culture supernatant was measured by using an ELISA kit (AssayMax Human Plasminogen activator inhibitor-1 ELISA Kit™, AssayPro LLC, St. Charles, MO, USA).

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay

Journal: The American Journal of Pathology

Article Title: A Protein C Deficiency Exacerbates Inflammatory and Hypotensive Responses in Mice During Polymicrobial Sepsis in a Cecal Ligation and Puncture Model

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Figure Lengend Snippet: Plasma coagulation factor assays of WT and PC+/− mice after CLP surgery. a: Plasma PC levels. The plasma PC levels were determined by ELISA before (WT, n = 4; PC+/−, n = 5), and 6 hours (WT, n = 5; PC+/−, n = 7) and 24 hours (WT, n = 5; PC+/−, n = 5) after CLP surgery in WT and PC+/− male mice. *, P < 0.001; #, P < 0.01. b: Plasma fibrinogen levels were determined by a coagulometric assay at 6 hours (WT, n = 4; PC+/−, n = 5) and 24 hours (WT, n = 15; PC+/−, n = 14) after, CLP surgery and in resting (WT, n = 11; PC+/−, n = 9) WT and PC+/− male mice. *, P = 0.016. c: Plasma PAI-1 levels were determined by ELISA before (WT, n = 3; PC+/−, n = 4), and 6 hours (WT, n = 3; PC+/−, n = 7) and 24 hours (WT, n = 5; PC+/−, n = 4) after, CLP surgery in WT and PC+/− male mice. d: Plasma FXII levels relative to the resting WT group, as measured by a coagulometric assay, before (WT, n = 4; PC+/−, n = 3) and 24 hours after (WT, n = 5; PC+/−, n = 3), CLP surgery. *, P = 0.05 (Kruskal-Wallis). e: Plasma FXI levels relative to the resting WT group as measured by a coagulometric assay, before (WT, n = 7; PC+/−, n = 5) and 24 hours after (WT, n = 5; PC+/−, n = 3), CLP surgery. No significant differences were observed.

Article Snippet: After four washes with PBS/Tween-20, 100 μl of 4 μg/ml rabbit anti-mouse PAI-1 polyclonal antibody (Molecular Innovations) in PBS was added to the wells and incubated for 2 hours.

Techniques: Coagulation, Enzyme-linked Immunosorbent Assay

Journal: The American Journal of Pathology

Article Title: A Protein C Deficiency Exacerbates Inflammatory and Hypotensive Responses in Mice During Polymicrobial Sepsis in a Cecal Ligation and Puncture Model

doi:

Figure Lengend Snippet: Gene expression ratio to RPL-19 of PAI-1 (a), MPO (b), IL-1β (c), and IL-6 (d) relative to the WT resting group (n = 3) in liver, kidney, and lung, as determined by RT-PCR in WT mice 24 hours after CLP (n = 3), PC+/− resting mice (n = 3), and PC+/− mice 24 hours after CLP (n = 4). a: In liver, PC+/− mice 24 hours after CLP were different from PC+/− resting and WT resting groups (P = 0.025, K-W), in kidney, differences between groups were not significant (K-W, multicomparison test). b: In liver, PC+/− resting mice were different from PC+/− mice 24 hours after CLP and WT resting groups (P = 0.055); in kidney, PC+/− mice 24 hours after CLP were different from all of the other groups (P = 0.02, K-W); in lung, PC+/− resting mice were different from PC+/− and WT mice 24 hours after CLP (P = 0.08, K-W). c: In liver, no significant differences between groups were found; in kidney: PC+/− group 24 hours after CLP was different from WT and PC+/− resting groups (P = 0.04, K-W); in lung: the PC+/− group after CLP was not different form the PC+/− resting group (T-K); P = not significant (K-W). d: In liver, PC+/− mice 24 hours after CLP were different from the WT resting group (P = 0.11, K-W); in kidney, PC+/− mice 24 hours after CLP were different from the WT and PC+/− resting groups (P = 0.032, K-W); in lung: no significant differences between the groups were found (K-W).

Article Snippet: After four washes with PBS/Tween-20, 100 μl of 4 μg/ml rabbit anti-mouse PAI-1 polyclonal antibody (Molecular Innovations) in PBS was added to the wells and incubated for 2 hours.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction