compound 1 pac 1  (MedChemExpress)

Journal: Frontiers in Microbiology

Article Title: Enterovirus A71 2B Inhibits Interferon-Activated JAK/STAT Signaling by Inducing Caspase-3-Dependent Karyopherin-α1 Degradation

doi: 10.3389/fmicb.2021.762869

Figure Lengend Snippet: Viral 2B induced KPNA1 degradation in various cell types and intrinsic apoptosis. (A,B) Degradation of KPNA1 and cleaved-caspase-3 were detected in HT-29 (A) and RD (B) cells. Cells transfected with plasmids expressing HA-tagged 2B and cell lysates were prepared 36 h post transfection with varying amounts of plasmids for western blot analyses with antibodies for KPNA1, HA, cleaved-caspase-3, or β-Tubulin. (C) 2B induced PARP cleavage as well as KPNA1 degradation. HeLa cells were treated with Cisplatin (30 μM) or PAC1 (50 μM), or transfected with different amounts of plasmids expressing HA-tagged 2B for 36 h. Whole cell lysates were collected for western blot analyses with specific antibodies. Ctl, untreated group; Cis, Cisplatin; PAC, PAC1. (D) Cell viabilities with 2B expression. HeLa cells were treated with Cisplatin or PAC1, or transfected with plasmids expressing HA-tagged 2B. The cells were assessed with the CCK-8 assay for cell viability. The values represented mean ± SD from triplicates of independent experiments and the viability of negative control cells without any treatment was set as 100%.

Article Snippet: Pro-caspase activating compound 1 (PAC1), a caspase-3 agonist, was ordered from Tocris Bioscience (Bristol, United Kingdom).

Techniques: Transfection, Expressing, Western Blot, CCK-8 Assay, Negative Control