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Control and blocking experiments were carried out in the same animal on 2 consecutive days.
Thus, stress fiber assembly and EPM receptor (β1 integrin) blocking experiments were carried out to explore their roles in causing EPM to affect MO.
Blocking experiments were carried out with Src family-selective tyrosine kinase inhibitors, PP1, PP2 (Biomol International, LP, of Plymouth Meeting, PA, USA) and PP3 (the inactive analogue, Calbiochem), at an end concentration of 10 μ M in line with a large body of literature.
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Receptor blocking experiment was carried out to confirm receptor-mediated endocytosis of Tf-C-SLN.
To detect the specific binding for α7 nAChRs, a blocking experiment was carried out using the selective α7 nAChR agonists (SSR180711, unlabeled A-582941 or A-844606), and the selective α4β2 nAChR agonist A-85380 (Tables 3, 4, 5, and 6).
All experiments were carried out in duplicate.
Receptor-blocking experiments were carried out using propranolol (5 mg/kg).
Antibody-blocking experiments were carried out with the same protocol, this time using PCSK9 at 2.5 μg/ml and incorporating a pre-incubation step at 4 °C for 1 h before cell treatment with various concentrations (0 100 μg/ml) of antibody.
In addition, comparative computational fluid dynamics (CFDs) analysis for the multi-block experiment was carried out to supplement the experimental limit and to analyze the local flow characteristics in detail.
The experiments were carried out in a randomized block design with three repetitions.
Experiments were carried out in triplicate for each time point (control and blocking).
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