Journal: Journal of Virology

Article Title: The Epstein-Barr Virus (EBV)-Induced Tumor Suppressor MicroRNA MiR-34a Is Growth Promoting in EBV-Infected B Cells

doi: 10.1128/JVI.07056-11

Figure Lengend Snippet: Canonical miR-34a growth control targets are poorly expressed in LCLs relative to HCT-116 cells. (a) Relative mRNA levels as determined by qRT-PCR of cyclin D1, D2, and E2; cdk4; cdk6; and c-Met in LCLs and HCT-116 cells. The asterisk indicates a value below the threshold for detection. (b) Western analysis of miR-34a targets in HCT-116 cells and EF3D LCLs expressing pcDNA3 vector (−) or pCDNA3-miR-34a (+). Quantitation of protein expression is noted below each band.

Article Snippet: The antibodies used were cyclin D1 and D2 (BD; 554203), cyclin E2 (Cell Signaling; 4132), cdk4 (Santa Cruz; H-22), cdk6 (Santa Cruz; C-21), and Met (Cell Signaling; 4560).

Techniques: Control, Quantitative RT-PCR, Western Blot, Expressing, Plasmid Preparation, Quantitation Assay

Journal: Nature microbiology

Article Title: N4BP1 restricts HIV-1 and its inactivation by MALT1 promotes viral reactivation.

doi: 10.1038/s41564-019-0460-3

Figure Lengend Snippet: Fig. 2 | N4BP1 is upregulated following IFN stimulation and restricts viral replication in T cells. a, N4BP1 mRNA levels were measured by RT–qPCR in Jurkat cells treated with IFN-α from human leukocytes (1,000 U ml−1) for the indicated periods of time. The data are shown as the mean ± s.d. of biological replicates (n = 3). b, Immunoblot analysis of N4BP1 in cell lysates from Jurkat cells treated with IFN-α for 48 h. β-actin was used as the loading control. c, N4BP1 mRNA levels were measured by RT–qPCR in primary human CD4+ T cells treated with IFN-α for 24 h. Individual points and means ± s.d. are shown (n = 3). d, Expression levels of IFNB1, N4BP1, BST-2 and ISG15 mRNAs were measured by qPCR in Jurkat cells infected with HIV-1 NL4-3 (MOI 0.01). The data are shown as the mean ± s.d. of biological replicates (n = 3). e, N4BP1 mRNA levels in spleens from humanized mice 6 weeks after HIV-1 infection (n = 8) or mock treatment (n = 6). Individual points and means ± s.d. are shown. f, Immunoblot analysis of N4BP1 in cell lysates from control and N4BP1-KO Jurkat cells (left panel). N.S., non-specific; IB, immunoblot. Replication of HIV-1 NL4-3 (MOI 0.01) in N4BP1-KO or control Jurkat cell lines (right panel). Infectivity of HIV-1 in the culture supernatants was measured by TZM-bl assay. g, Immunoblot analysis of N4BP1 in cell lysates from control cells and N4BP1-KO Jurkat cells inducibly reconstituted with N4BP1 by using the Tet-on system and Dox stimulation for 24 h (left panel). Replication of HIV-1 NL4-3 (MOI 0.01) in control or N4BP1-KO Jurkat cell lines reconstituted with or without N4BP1 by Dox treatment (right panel). The infectivity of HIV-1 in the culture supernatants was measured by TZM-bl assay. h, Immunoblot analysis of Jurkat cells stably expressing FLAG-tagged N4BP1 (left panel). Replication of HIV-1 NL4-3 (MOI 0.01 or 0.001) in FLAG–N4BP1-expressing or control Jurkat cell lines (right panel). The infectivity of HIV-1 in the culture supernatants was measured by TZM-bl assay. P values were calculated using unpaired two-tailed Student’s t-test. *P < 0.05; **P < 0.01; ***P < 0.005.

Article Snippet: Proteins were labelled with antibodies against HIV-1 Env (1:2,000; 16H3, catalogue no. 12559, NIH AIDS Research and Reference Reagent programme), p24 (1:2,000, polyclonal; ViroStat), Vif (1:2,000; no. 319; NIH AIDS Research and Reference Reagent programme), Nef (1:2,000; 3D12, ThermoFisher), Vpr (1:2,000; 8D1, Cosmo Bio), Vpu (1:2,000; no. 969, NIH AIDS Research and Reference Reagent programme), Flag (1:1,000; monoclonal; F7425, Sigma, or polyclonal; F7425, Sigma), mouse IgG isotype control (1:1,000; no. 31903, ThermoFisher), β-actin (1:1,000; polyclonal; sc-1615, Santa Cruz), MALT1 (1:1,000; no. 2494, Cell Signaling Technology), GAPDH (1:1,000; sc-47724, Santa Cruz), α-tubulin (1:1,000; T9026, Sigma), IκBα (1:1,000; C-21, Santa Cruz), GFP (1:1,000; ab290, Abcam) and N4BP1 (1:2,000, ab197079, Abcam).

Techniques: Quantitative RT-PCR, Western Blot, Control, Expressing, Infection, Stable Transfection, Two Tailed Test

Journal: Toxins

Article Title: Host Genotype and Weather Effects on Fusarium Head Blight Severity and Mycotoxin Load in Spring Barley

doi: 10.3390/toxins14020125

Figure Lengend Snippet: Genotype ranking according to detected Fusarium toxin contents in mature barley heads after bruised grain inoculation in the seasons 2018 to 2020. The heat map represents scored mean ranks according to detected contents of DON, NIV, DON3G, 3ADON, enniatins A, A1, B and B1 in mature barley heads in inoculated sub plots. Mean ranks of individual genotypes and the respective standard error of the mean are presented in the grey shaded box. Low ranks indicate high FHB resistance; high ranks indicate low FHB resistance. Error bars indicate standard error of the mean.

Article Snippet: Unlabelled reference compounds (deoxynivalenol-3-glucoside (D3G), 15-acetyl-deoxynivalenol (15-ADON), T2-toxin) and some labelled standards ( 13 [C 15 ]-DON, 13 [C 17 ]-3-ADON, 13 [C 22 ]-HT2-toxin and 13 [C 21 ]-D3G) were purchased from Biopure (Tulln, Austria).

Techniques: