Additional file 1 of Indigofera suffruticosa aerial parts extract induce G2/M arrest and ATR/CHK1 pathway in Jurkat cells

Additional file 1: Supplementary Table S1. Antibodies used in this study. Supplementary Fig. S1. Representative plot of Annexin V staining results. This figure is related to Fig. 2D. Supplementary Fig. S2. The caspase-3/7 activities in Jurkat cells after 12 h of ISAE treatment were assessed by Caspase-Glo® 3/7 assay (Promega) according to the manufacturer's instructions. The graph was expressed as fold changes to control group. Supplementary Fig. S3. Relative quantitative analysis of tryptanthrin, indigo, and indirubin in ISAE. The selected ion current chromatograms of the ISAE extract, tryptanthrin, indigo, and indirubin were carried out through MS full scan experiment in positive mode. The relative abundance of the three selected compounds in ISAE extract were calculated based on comparing the peak area ratios with standard compounds. Supplementary Fig. S4. Cell gating for cell cycle analysis. For cell cycle analysis, cells were gated using forward scatter (FSC) and side scatter (SSC) properties. This helped in excluding cell debris and selecting the desired cells (within the black circle) for analysis. The gating area was established based on the solvent control group (0 μg/mL of ISAE) and was consistently applied to all other groups. This illustration corresponds to Fig. 2A. Original images of western blot. Multiple exposure images of WB

Chemical Constituents and Bioactive Principles from the Mexican Truffle and Fermented Products of the Derived Fungus Ustilago maydis MZ496986

To look in-depth into the traditional Mexican truffle, this study investigated the phytochemical and pharmacological properties of field-collected corn galls and the fermentate of its pathogen Ustilago maydis MZ496986. Here, we established the chemical profiles of both materials via the gradient HPLC-UV method and successfully identified six previously unreported chemical entities, ustilagols A–F (1–6), and 17 known components. Compounds 3, 5, and 9 exhibited potent nitric oxide production inhibitory activities in murine brain microglial BV-2 cells (IC50 = 6.7 ± 0.5, 5.8 ± 0.9, and 3.9 ± 0.1 μM) without cytotoxic effects. DIMBOA (9) also attenuates lipopolysaccharide (LPS)-stimulated NF-κB activation in RAW 264.7 macrophages (IC50 = 58.1 ± 7.2 μM). Ustilagol G (7) showed potent antiplatelet aggregation in U46619-stimulated human platelets (IC50 = 16.5 ± 5.3 μM). These findings highlighted the potential of corn galls and U. maydis MZ496986 fermentate as functional foods for improving inflammation-related discomforts and vascular obstruction

Characterization of the effects of a 7 amino-acid deletion in p6^gag to the HIV-1 proteins expression, release and maturation.

<p>MT2 cells were infected with wild type (wt) or deleted-type (7d) recombinant viruses. After 12, 24, 36 and 48 hours, supernatant was collected and pelleted by ultracentrifugation. (A) Western blot analysis of the cell lysates (left panel) and viral lysates (right panel) from cells infected with wt or 7d viruses. (B) The relative expression levels of PR and RT in the viral lysates of cells infected with wt or 7d virus. The total arbitrary densitometer units of PR and RT were standardized by p24 and normalized to those of wt in parallel experiments. The images were analyzed with Image J software. (C) The ratios of p24 vs. Pr55 (maturation index) in the viral lysates at different time points after infection were calculated. The total arbitrary densitometer units of each hours post infection were normalized to those of wt in parallel experiments. All results were representative of two independent experiments. (D) Electron microscopic (EM) examination of the viral particles of cells infected with wt or 7d recombinant viruses. MAGIC-5 cells were fixed and processed for transmission EM at different time points after they were infected with wt or 7d viruses. Scale bar indicates 200 nm. (E) Quantification of relative proportions of mature vs. immature virions released at different time points in the cells infected with wt or 7d viruses using EM. The method of virion quantification has been described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0114441#pone.0114441-Fujii1" target="_blank">[43]</a>.</p

Comparisons of the changes of CD4 cell count and HIV-1 viral loads after the first clinical visit among the following four groups of treatment naïve patients.

<p>Men who have sex with men (MSM, 357 patients for CD4 cell count and 371 patients for viral loads analysis) vs. injection drug users (IDUs, 129 patients for CD4 cell count and 128 patients for viral loads analysis) (Fig. 1A and 1B); patients infected with CRF07_BC vs. infected with subtype B (Figs. 1C and 1D). A generalized estimating equation model was used for the analyses.</p

V3 amino acid sequences and predicted phenotypes of different HIV-1 isolates in Taiwan.

<p>A dash indicated a deletion or lack of an insertion.</p>a<p>The phenotype prediction based on 2 amino acid insertion/deletion between position 14 and 15, as well as variable amino acid positions (11, 18, 19, 23, 24 and 25) of V3 regions.</p>b<p>Geno2pheno (<a href="http://coreceptor.bioinf.mpi-inf.mpg.de/index.php" target="_blank">http://coreceptor.bioinf.mpi-inf.mpg.de/index.php</a>), false-positive rate of 0.01.</p>c<p>Position-Specific Scoring Matrix (PSSM) (<a href="http://indra.mullins.microbiol.washington.edu/webpssm/" target="_blank">http://indra.mullins.microbiol.washington.edu/webpssm/</a>).</p><p>V3 amino acid sequences and predicted phenotypes of different HIV-1 isolates in Taiwan.</p

Demographic data, risk factors and clinical characteristics of HIV-1-infected patients recruited in the study.

<p>Demographic data, risk factors and clinical characteristics of HIV-1-infected patients recruited in the study.</p

Univariate and multivariate generalized estimating equations models of factors associated with CD4 cell counts.

a<p>The mode of transmission was removed from multivariate model II.</p><p>Univariate and multivariate generalized estimating equations models of factors associated with CD4 cell counts.</p

Univariate and multivariate generalized estimating equations model of factors associated with HIV-1 viral loads.

a<p>The mode of transmission was removed from multivariate model II.</p><p>Univariate and multivariate generalized estimating equations model of factors associated with HIV-1 viral loads.</p

Distribution of HIV-1 subtypes and circulating recombinant forms (CRFs) in different groups of patients recruited in this cohort study.

a<p>Includes 4 heterosexual female infected with subtype B, 3 heterosexual female infected with CRF01_AE, and 3 IDUs infected with CRF07_BC.</p><p>Distribution of HIV-1 subtypes and circulating recombinant forms (CRFs) in different groups of patients recruited in this cohort study.</p

Comparison of the replication kinetics of different HIV-1 isolates from patients infected with CRF07_BC or subtype B (A and B) as well as recombinant HIV-1 virus with or without a 7 amino-acid deletion in the p6^gag protein.

<p>(A) PBMCs were infected with fixed amounts of different HIV-1 subtype B (square) or CRF07_BC (circle) isolates and cultural supernatants were collected at days 4, 7, 11, 14, 18 and 21 post-infection. (B) The representative replicative curves of CRF07_BC and subtype B isolates deduced from Fig. 2A. (C) MT2 cells were infected with wild type (wt) or recombinant mutant virus with a 7 amino-acid deletion at the p6^gag (7d). Supernatants were collected at days 2, 4, 6, 8, 10, 12, and 14 post-infection. Viral replication was monitored through p24 antigen production. One-way analysis of variance (ANOVA) and Tukey's post hoc test were used to estimate the differences between subtypes, or between the recombinant viruses.</p