Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Scutellarein selectively targets multiple myeloma cells by increasing mitochondrial superoxide production and activating intrinsic apoptosis pathway.

doi: 10.1016/j.biopha.2018.09.024

Figure Lengend Snippet: Fig. 2. Scutellarein treatment activated the intrinsic apoptosis pathway in multiple myeloma cells in vitro. A–C, analysis of caspase-9, -8 and -3 activities in IM-9, MM.1R or viable circulating B lymphocytes (CBL) after treatment with indicated concentrations of scutellarein for 24 h. Caspase activity was measured by the relative fluorescent units (RFU) of cleaved substrates. Data was normalized to the vehicle-treated CBL group. D–I, influence of FADD or APAF1 knockdown on the apoptosis- inducing effect of 400 μg/ml scutellarein treatment for 24 h. shRNA targeting green fluorescent protein gene (sh-NC) was used as negative control for FADD or APAF1 knockdown. Data in DeI were normalized to the un-transfected group (WT) in each cell line. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Article Snippet: Antibodies for detecting FADD (PA1464, BosterBio, Pleasanton, USA), Apaf-1 (PA1249-1, BosterBio), active Caspase-3 (269518, Novus Biologicals, Littleton, USA) Bax (AF820, R& D Systems, Minneapolis, USA), Bcl-2 (RP1003, BosterBio) and GAPDH (HPA040067, Atlas Antibodies, Bromma, Sweden) were applied following manufacturer’s instructions.

Techniques: In Vitro, Activity Assay, Knockdown, shRNA, Negative Control, Transfection

Journal: PLoS ONE

Article Title: TRAIL Receptor Signaling Regulation of Chemosensitivity In Vivo but Not In Vitro

doi: 10.1371/journal.pone.0014527

Figure Lengend Snippet: In panel A, isogenic BJAB cells expressing GFP or GFP-FADD-DD and FADD-DDV108E were treated with increasing doses of Fas ligand, TRAIL, or agonistic anti-DR4 and anti-DR5 antibodies. The differences in responses indicate that both FADD-DD expressing cell lines were resistant to all TRAIL R targeted drugs, but that only FADD-DD expressing cells are resistant to FasL. In panel B, DISC IP experiments precipitating Fas or DR5 followed by western blotting for casp-8 or FADD demonstrate an increased recruitment of FADD-DD in place of endogenous FADD to the receptors. Panel C, illustrates that on activation of other signaling pathways leading to IkB degradation, and JNK phosphorylation, FADD-DD blocks signaling. This block leads to these events for both FasL and TRAIL, and that FADD-DD V108E blocks signaling only for TRAIL.

Article Snippet: Blots were blocked with 5% milk in TBST and incubated with antibodies that recognize IKappaB-alpha, phospho-JNK, JNK, Caspase 8, Caspase 3 (Cell Signaling Technologies, Danvers, MA), FADD (BD Biosciences, Franklin Lakes, NJ).

Techniques: Expressing, Western Blot, Activation Assay, Blocking Assay