Journal: bioRxiv

Article Title: TNF controls a speed-accuracy tradeoff in the apoptotic decision to restrict viral spread

doi: 10.1101/2020.02.20.958942

Figure Lengend Snippet: A , Quantification of cell death after treatment with TNF or TNF supplemented with Actinomycin D. B , Representative images of cells assayed for DISC assembly by Traf2-Caspase 8 promixity ligation. C ,Quantification of number of DISC spots per cell for untreated and TNF-treated cells. Line shows the gate selected to distinguish positive cells. Number of cells = 475(Untreated), 261(TNF-tx) D , Quantification of TNF-induced ciap1 and 2 by qPCR. E , Quantification of cell death for cells treated with 0 or 0.3ng/ml TNF and vehicle or 50pg/ml LCL-161. F , Quantification of cell death for cells treated constitutively or overnight with 10ng/ml TNF. All scale bars=100μm. D-F Data are mean +/− s.d. All scale bars,100μm.

Article Snippet: 12.5×10 4 3T3 cells were seeded per well of black 96-well plates in imaging media (Fluorobrite with 10% NBCS, 2mM L-glutamine, 10U/ml Penicillin and 10μg/ml Streptomycin) supplemented, where indicated, with dose titrations of recombinant mouse TNFα (Cell Signaling Technologies), 30μM Z-VAD(OMe)-FMK (Cayman Chemical), 30μM Necrostatin-1 (Sigma Aldrich), 0.25μg/ml Actinomycin D (ThermoFisher), or the specified doses of LCL-161 (Cayman Chemical).

Techniques: Ligation

Journal: bioRxiv

Article Title: TNF controls a speed-accuracy tradeoff in the apoptotic decision to restrict viral spread

doi: 10.1101/2020.02.20.958942

Figure Lengend Snippet: A , Quantification of infected cells after treatment with dose titrations of TNF supplemented with vehicle or 30μM Z-Vad and 30μM Necrostatin-1. Cells were infected with MOI 1 of HSV-1. B , Quantification of infected cells after treatment with 0 or 3ng/ml TNF supplemented with dose titrations of LCL-161. Cells were infected with MOI 1 of HSV-1. C , Average viral load per cell quantified from HSV-1 fluorescence after infection with MOI 10 HSV-1. D , Quantification of the infection rate for cells based on the viral load of their nearest neighbor. E , Death rates for cells treated with dose titrations of TNF and infected with MOI 10 of HSV-1 or left uninfected. F , Quantification of the coupling between infected and uninfected cell death rates. A-C, E-F, Data are mean +/− s.d. D , Data are mean+/− s.e.m.

Article Snippet: 12.5×10 4 3T3 cells were seeded per well of black 96-well plates in imaging media (Fluorobrite with 10% NBCS, 2mM L-glutamine, 10U/ml Penicillin and 10μg/ml Streptomycin) supplemented, where indicated, with dose titrations of recombinant mouse TNFα (Cell Signaling Technologies), 30μM Z-VAD(OMe)-FMK (Cayman Chemical), 30μM Necrostatin-1 (Sigma Aldrich), 0.25μg/ml Actinomycin D (ThermoFisher), or the specified doses of LCL-161 (Cayman Chemical).

Techniques: Infection, Fluorescence