Journal: Biochimica et biophysica acta

Article Title: Increases in intracellular calcium perturb blood-brain barrier via protein kinase C-alpha and apoptosis.

doi: 10.1016/j.bbadis.2015.10.016

Figure Lengend Snippet: Fig. 4. Amiloride and apocynin reduce PKC activity and uPA activity affected by PKC-α. HBMEC were exposed to 4 h OGD (OGD) with or without reperfusion (R) in the absence or presence of inhibitors for uPA (amiloride), NADPH oxidase (apocynin), intracellular calcium (BAPTA-AM) or PKC-α (RO-32-0432). Amiloride and apocynin decreased the rise in total PKC (A) and PKC-α (B) activities seen during OGD ± R. tPA activity is increased during OGD ± R and treatment with BAPT-AM and RO-32-0432 show no effect (C). uPA activity is also increased during OGD ± R and BAPTA-AM and RO-32-0432 normalised the increases seen (D). Data represented as mean ± SEM from n ≥3.* P b 0.05 compared to Normoxia. † P b 0.05 compared to re- spective untreated group.

Article Snippet: PAI-1-bound active uPA or tPA was detected by anti-tPA or anti-uPA primary antibodies (1:50, Santa Cruz Biotechnology) and primary antibodies were probed with horseradish peroxidase-linked secondary antibodies (1:5000, Santa Cruz).

Techniques: Activity Assay

Journal: Pharmaceutical research

Article Title: A Role for Low Density Lipoprotein Receptor-related Protein 1 in the Cellular Uptake of Tissue Plasminogen Activator in the Lungs

doi: 10.1007/s11095-015-1763-6

Figure Lengend Snippet: The concentration of intracellular fraction mtPA was lower over time in the presence of LRP1 inhibitor, receptor associated protein (RAP) in primary liver and lung cell suspensions (20,000 cells each). Cells were exposed to mtPA (50ng) at time 0 in the presence or absence of RAP (400nM). Representative Western blots of intracellular fraction mtPA over time in (A) primary whole liver cell lysates and (B) primary whole lung cell lysates. These findings were corroborated by the measurement of intracellular fraction mtPA concentrations measured by ELISA in (C) liver cell (*p=0.0132 at 1 min) and (D) lung cell (*p=0.009; +p<0.001; #p=0.066) lysates as well as in extracellular fraction (supernatant) media of (E) liver (*p=0.005; +p=0.0005) and (F) lung (*p=0.024; +p=0.001) cell suspensions. Data are the mean (±S.E.M.) of 3-4 samples/group and p values were generated from post ANOVA Holm-Sidak tests.

Article Snippet: Measurement of mtPA Uptake by Lung and Liver Cell Suspensions The concentration of mtPA in lung and liver cell lysates (intracellular fraction) and supernatants (extracellular fraction) was detected using mtPA total antigen assay ELISA kit (MTPAKT-TOT, Molecular Innovations).

Techniques: Concentration Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Generated