Journal: EMBO Reports

Article Title: IFN‐stimulated metabolite transporter ENT3 facilitates viral genome release

doi: 10.15252/embr.202255286

Figure Lengend Snippet: BMDMs were treated with different MOIs of E. coli DH5α for 30 min and further cultured for various length of time as indicated. The dynamic of ENT3 expression was determined by measuring Slc29a3 transcripts via RT‐qPCR. Results from three biological replicates ( n = 3) are shown. The efficiency of phagocytosis inhibition by Cytochalasin D treatment in macrophages. BMDMs were incubated with pHrodo Red pre‐labeled GFP‐expressing E. coli BL21 (LiveEC) at MOI 100 in the presence or absence of phagocytosis inhibitor, Cytochalasin D (CytoD), for 30 min. Images were captured by Zeiss Axio Observer 7 40X lens after washing out the extracellular E. coli . Results were representative images from three biological replicates ( n = 3). Scale bars represent 20 μm. Unpaired two‐tailed Student's t ‐test was used for statistical analysis, ** P < 0.01, ***P < 0.001, n.s. no significance. Data are shown as mean ± SEM.

Article Snippet: Escherichia coli DH5α (Protech) were transfected with pUC19 plasmid which has ampicillin resistance gene as a selection marker.

Techniques: Cell Culture, Expressing, Quantitative RT-PCR, Inhibition, Incubation, Labeling, Two Tailed Test

Journal: EMBO Reports

Article Title: IFN‐stimulated metabolite transporter ENT3 facilitates viral genome release

doi: 10.15252/embr.202255286

Figure Lengend Snippet: BMDMs were treated with E. coli (MOI = 100) for 30 min and further cultured for various length of time as indicated. The dynamic of ENT3 expression was determined by measuring Slc29a3 transcripts via RT‐qPCR. Phagocytosis inhibitor, Cytochalasin D (CytoD), pretreated BMDMs were incubated with E. coli (MOI = 100) and subjected for Slc29a3 expression analysis via RT‐qPCR 6 h post‐infection. The bacterial effector molecules generated from different treatments. Live, heat‐killed (HK), gentamicin‐killed (GK) E. coli , or metabolite‐containing supernatant (Meta), were prepared. Different bacterial components were applied as stimulants to BMDMs. Slc29a3 expression was analyzed after 30 min of stimulation and a total of 6 h incubation. BMDMs were treated with different doses of LPS for 30 min, harvested 6 h post‐stimulation, and subjected for RT‐qPCR analysis. Data information: The expression level was calculated relative to Rpl19 and normalized to untreated group as 1. The results shown are combined from three biological replicates ( n = 3), unpaired two‐tailed Student's t ‐test was used for statistical analysis, * P < 0.05, ** P < 0.01, *** P < 0.001, n.s. no significance. Data are shown as mean ± SEM.

Article Snippet: Escherichia coli DH5α (Protech) were transfected with pUC19 plasmid which has ampicillin resistance gene as a selection marker.

Techniques: Cell Culture, Expressing, Quantitative RT-PCR, Incubation, Infection, Generated, Two Tailed Test

Journal: EMBO Reports

Article Title: IFN‐stimulated metabolite transporter ENT3 facilitates viral genome release

doi: 10.15252/embr.202255286

Figure Lengend Snippet: A The Schematic diagram of TLR4 signaling pathways. B BMDMs from WT or MyD88 −/− mice were treated with 10 ng/ml of LPS or live E. coli (MOI = 100) and subjected for Slc29a3 expression analysis. C, D (C) WT or (D) MyD88 −/− BMDMs were stimulated with 10 ng/ml LPS or live E. coli (MOI = 100) in the presence of 10 μM TBK inhibitor, MRT67307, and analyzed for Slc29a3 expression. E The 10 ng/ml LPS‐treated BMDMs were examined for Ifnb1 expression after 30 min of stimulation and a total of 6 h incubation. F BMDMs were treated with 100 U/ml of IFN‐β for various length of time and subjected for Slc29a3 expression. Data information: The expression level was calculated relative to Rpl19 and normalized to untreated group as 1. The results shown are combined from three biological replicates ( n = 3), unpaired two‐tailed Student's t ‐test was used for statistical analysis, * P < 0.05, ** P < 0.01, *** P < 0.001, n.s. no significance. Data are shown as mean ± SEM.

Article Snippet: Escherichia coli DH5α (Protech) were transfected with pUC19 plasmid which has ampicillin resistance gene as a selection marker.

Techniques: Protein-Protein interactions, Expressing, Incubation, Two Tailed Test

Journal: EMBO Reports

Article Title: IFN‐stimulated metabolite transporter ENT3 facilitates viral genome release

doi: 10.15252/embr.202255286

Figure Lengend Snippet: A The brief schematic diagram of type I IFN downstream signaling cascades. B–E BMDMs from WT, IFNAR1 −/− (B), or STAT1 −/− (C) mice were treated with 10 ng/ml LPS, live E. coli (MOI = 100), or 100 U/ml IFN‐β, harvested and examined for Slc29a3 induction. BMDMs from WT, IFNAR1 −/− (D) or STAT1 −/− (E) mice were treated with 100 U/ml IFN‐β, 50 μg/ml poly(I:C), or EMCV (MOI = 10), harvested and examined for Slc29a3 induction. F–H The expression dynamic of Slc29a3 was profiled in BMDMs treated with two different doses of IFN‐α (F), IFN‐β (G), IFN‐γ (H) for various length of time as indicated. Data information: The expression level was calculated relative to Rpl19 and normalized to untreated group as 1. The results shown in (B–E) are from three biological replicates ( n = 3) and in F to H from two biological replicates ( n = 2). Unpaired two‐tailed Student's t ‐test was used for statistical analysis, **P < 0.01, ***P < 0.001, n.s. no significance. Data are shown as mean ± SEM.

Article Snippet: Escherichia coli DH5α (Protech) were transfected with pUC19 plasmid which has ampicillin resistance gene as a selection marker.

Techniques: Expressing, Two Tailed Test

Journal: EMBO Reports

Article Title: IFN‐stimulated metabolite transporter ENT3 facilitates viral genome release

doi: 10.15252/embr.202255286

Figure Lengend Snippet:

Article Snippet: Escherichia coli DH5α (Protech) were transfected with pUC19 plasmid which has ampicillin resistance gene as a selection marker.

Techniques: Virus, Recombinant, shRNA, Sequencing, Transfection, SYBR Green Assay, Staining, Chromatin Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Imaging, Software

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Targeting the Nonmevalonate Pathway in Burkholderia cenocepacia Increases Susceptibility to Certain β-Lactam Antibiotics

doi: 10.1128/AAC.02607-17

Figure Lengend Snippet: Bacterial strains and plasmids

Article Snippet: DH5α λpir , λpir, for propagation of suicide plasmid pSC200 and its derivatives , Biomedal.

Techniques: Plasmid Preparation, Control, Transformation Assay

Journal: Molecular Cell

Article Title: Impact of chromatin context on Cas9-induced DNA double-strand break repair pathway balance

doi: 10.1016/j.molcel.2021.03.032

Figure Lengend Snippet:

Article Snippet: CloneCatcher DH5α electrocompetent E. coli , Genlantis , Cat# C810111.

Techniques: Western Blot, Virus, Recombinant, Transfection, Lysis, Viability Assay, Protease Inhibitor, Reverse Transcription, Plasmid Preparation, Software