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Maximal response with 5 µM MG132 was reached at 20 µM niclosamide (IC50 of ∼5 µM).
The 20 mg/kg SD-208 dose was chosen to achieve specific inhibition of the TβRI kinase, whereas the 60 mg/kg dose was chosen to achieve a maximal response with minimal inhibition of other pathways [19].
The data were normalized to Renilla null luciferase activity and expressed as a percentage of the maximal response with Dex before being plotted ± the standard error of the mean, unless otherwise noted.
To investigate whether stimulation of the maximal response with the 5 mix and DHT was the result of cell proliferation, we used a Renilla luciferase construct in our assay.
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The molecular fingerprints were obtained by weighting maximal responses with the respective antibody surface coverage and normalized to anti-CD63 responses.
As shown in Figure 5C, BK stimulated PKC-α translocation in a time-dependent manner with a maximal response within 5 min, which was attenuated by pretreatment with either BAPTA/AM (30 μM) or Gö6976 (1 μM).
We next determined whether the translocation of p47phox involved in BK-induced responses, as shown in Figure 3C, BK stimulated translocation of p47phox from the cytosol to the membrane with a maximal response within 3 min, which was attenuated by pretreatment with Apo.
As shown in Figure 7A (upper part), BK time-dependently stimulated binding of c-Fos and c-Jun to the MMP-9 promoter with a maximal response within 30 min, which was attenuated by pretreatment with BAPTA/AM, Gö6976, Apo, DPI, or NAC.
As shown in Figure 6A, ET-1 enhanced NF-κB transcriptional activity in a time-dependent manner with a maximal response within 60 min, which was significantly inhibited by pretreatment with an inhibitor of NF-κB (Bay11-7082, 10 nM).
Under our assay conditions (cells were in serum-free culture medium for 24 h) both caspase-9 and caspase-8 displayed a larger basal activity than caspase-3 when compared with the corresponding maximal response obtained with STS.
HEMADO retained an ability to activate ERK1/2 phosphorylation and to inhibit FSK-stimulated cAMP accumulation via A3 receptor W243F, albeit with a lower maximal response compared with the wild-type A3 receptor.
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