Journal: iScience

Article Title: Heat-induced SIRT1-mediated H4K16ac deacetylation impairs resection and SMARCAD1 recruitment to double strand breaks

doi: 10.1016/j.isci.2022.104142

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Article Snippet: Hela human cell line , ATCC , CRL-12401; RRID: CVCL_W341.

Techniques: Virus, Generated, Control, Recombinant, Protease Inhibitor, Extraction, cDNA Synthesis, Plasmid Preparation, Software

Journal: iScience

Article Title: Streptococcus agalactiae and Escherichia coli induce distinct effector γδ T cell responses during neonatal sepsis

doi: 10.1016/j.isci.2024.109669

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Article Snippet: Streptococcus agalactiae (Group B Streptococcus , GBS) COH-1 , American Type Culture Collection (ATCC) , COH-1, type III BAA-1176.

Techniques: Virus, Recombinant, Cell Stimulation, Enzyme-linked Immunosorbent Assay, Gene Expression, Software

Journal: Immunity

Article Title: Sensing microbial viability through bacterial RNA augments T follicular helper cell and antibody responses

doi: 10.1016/j.immuni.2018.02.015

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Recombinant mouse IFN-b , R&Dsystems , Cat#12401-1.

Techniques: Recombinant, Staining, Lysis, Cell Isolation, Software

Journal: Communications Biology

Article Title: Generation of transgenic mice expressing a FRET biosensor, SMART, that responds to necroptosis

doi: 10.1038/s42003-022-04300-0

Figure Lengend Snippet: a , b primary MEFs (pMEFs) ( a ) and immortalized MEFs (iMEFs) ( b ) were untreated or stimulated with TBZ (10 ng/mL TNF, 0.5 μM BV6, 20 μM zVAD) for 12 h. MEFs were generated from SMART Tg mice, and MEFs with less than ten passages were used as pMEFs. Cell death was determined with the LDH release assay. Results are mean ± SD of triplicate samples. c Western blots of lysates from untreated iMEFs from Mlkl −/− or wild-type mice, and untreated or IFNβ (2000 IU/mL)-treated pMEFs from SMART Tg mice. Blots were probed with anti-MLKL and anti-actin antibodies. d pMEFs were pretreated with IFNβ, and then stimulated with TBZ for 4 h. Cell viability was determined with the LDH release assay. Results are mean ± SD of triplicate samples. e – h pMEFs were untreated or stimulated with the indicated combination of agents (10 ng/mL TNF, 0.5 μM BV6, 20 μM zVAD, 5 μM GSK’872), and the FRET/CFP ratio was calculated. Pseudocolored images show cellular changes in the FRET/CFP ratio values in response to the indicated stimulations ( e , g ). FRET/CFP responses are color-coded according to the color scales (right). White arrowheads and white arrows indicate cells undergoing necroptosis ( e ) and apoptosis ( g ), respectively. Scale bars, 20 μm. Maximum changes of the FRET/CFP ratio are shown ( f , h ). Results mean ± SE ( n = 10 cells per condition). Each dot indicates an individual cell. Statistical analyses were performed by the unpaired two-tailed Student’s t -test ( a , b , d , h ) or one-way ANOVA with Tukey’s multiple comparison test ( f ). All results are representative of at least two independent experiments.

Article Snippet: We purchased the following reagents as follows: BV6 (B4653, ApexBio), cisplatin (AG-CR1-3590, Adipogen), GSK’872 (530389, Merck), IFNβ (12401-1, PBL-Assay Science), LPS (434, List Labs), nigericin (AG-CN2-0020, Adipogen), Murine TNF (PMC3013, Thermo Fisher Scientific), zVAD (3188-v, Peptide Institute), and 5z7Ox (499610, Calbiochem).

Techniques: Generated, Lactate Dehydrogenase Assay, Western Blot, Two Tailed Test

Journal: Viruses

Article Title: TNF Signaling Dictates Myeloid and Non-Myeloid Cell Crosstalk to Execute MCMV-Induced Extrinsic Apoptosis

doi: 10.3390/v12111221

Figure Lengend Snippet: TNF drives ∆M36-dependent apoptosis in endothelial cells and fibroblasts. ( A ). Relative viability of WT BMDM and SVEC4-10 cells infected with K181 or ∆M36 (MOI = 10) for 24 h in the absence or presence of murine TNF (25 ng/mL). TNF was added to medium 1 hpi. Graph reflects data pooled from three independent experiments, each containing 3 replicates. ( B ). IB showing the appearance of Cl-C8 and Cl-C3 with loading control β-actin 14 hpi in WT or SVEC4-10 cells under indicated conditions. Data are representative of 2 independent experiments. ( C – E ). Relative viability of SVEC4-10 treated with no cytokine, murine IFNβ or IFNγ (100 ng/mL each), ( C ), MEFs of indicated genotypes treated with murine TNF (25 ng/mL) ( D ) or HF treated with human TNF (100 ng/mL) ( E ) and infected with K181 or ∆M36 (MOI = 10) at 24 hpi. All cytokines were added to the medium at 1 hpi with virus. C-E reflect data pooled from two independent experiments, each containing 3 replicates. Lines above bars indicate the two groups compared for significance values using paired, parametric Student’s t -test where * is <0.05, ** is <0.01, and n.s. represents nonsignificant.

Article Snippet: All cells were maintained at 37 °C in a humidified 5% CO 2 incubator. zVAD-FMK (SM001) was from SM Biochemical, Anaheim, CA, USA; murine TNF (315-01A-20UG) and human TNF (300-01A) were from PeproTech, Cranbury, NJ, USA; and murine IFNβ (12401-1) and IFNγ (12500-2) were from PBL Assay Science, Piscataway, NJ, USA.

Techniques: Infection