Supplementary Figures from 3-hydroxyisobutyryl-CoA hydrolase involved in isoleucine catabolism regulates triacylglycerol accumulation in <i>Phaeodactylum tricornutum</i>

Supplemental Figure 1. Sequence alignment of the human PCC (HsPCC_PCC1), Ruegeria pomeroyi PCC (RpPCC_PCC1) a subunits, and the predicated PCC a subunits of Phaeodactylum tricornutum (PtPCC_PCC1) and Thalassiosira pseudonana (Tp_PCC1). Residues in the active site are indicated by arrows. Supplemental Figure 2. Sequence alignment of the human PCC (HsPCC_PCC2), Ruegeria pomeroyi PCC (RpPCC_PCC2) b subunits, and the predicated PCC b subunits of Phaeodactylum tricornutum (PtPCC_PCC2) and Thalassiosira pseudonana (Tp_PCC2). Supplemental Figure 3. Alignment of the amino acid sequences HIBCH from Homo sapiens (HsHIBCH), Arabidopsis thaliana (AtCHY1), Saccharomyces cerevisiae (ScHIBCH) and the predicted HIBCH in Chlamydomonas reinhardtii (CrHIBCH) and Phaeodactylum tricornutum (PtHIBCH). Amino acid residues identical in five, four, and three of the sequences are in red, green and blue respectively. Cleavage sites of mitochondria transit predicted by TargetP and the calculated score mTP are respectively shown by arrows and the numbers below the sequences. C-terminal peroxisome targeting signal are boxed. Asterisks above the sequence indicate proposed catalytic residues. Supplemental Figure 4. Phylogenetic relationship of the HIBCH protein family. The ML tree was generated using the MrBayes program. HIBCH are grouped into three distinct clades. Numbers above branches represent the support values (Bayesian posterior probabilities). The scale bar represents the number of nucleotide replacements per site. Supplemental Figure 5. Principal component analysis (PCA) of metabolic profiles in wild-type (WT), two HIBCH overexpression lines (hibch-OE1 and hibch-OE3) and one PCC RNAi silenced line (pcc1p). (a) Scores plot shows separation of the samples among the four groups; (b) loading plot indicated the main metabolites contributing to the separation

Supplementary Data Sets from 3-hydroxyisobutyryl-CoA hydrolase involved in isoleucine catabolism regulates triacylglycerol accumulation in <i>Phaeodactylum tricornutum</i>

Supplemental Data S1: MCEE gene distribution in different organisms. Supplemental Data S2: Primers used in this study for qPCR and the construction of RNAi expression vector, overexpression vector and GFP fusion vector. Supplemental Data S3: Metabolite contents (mg g-1 dry biomass) in wild-type, two overexpression lines (hibch-OE1 and hibch-OE3) and one silenced strain (pcc1p). Values are presented as the mean of three biological replicates. Supplemental Data S4: The values of fold change in transcript levels of genes encoding componens involved in BCAA degradation

Supplementary Figures from 3-hydroxyisobutyryl-CoA hydrolase involved in isoleucine catabolism regulates triacylglycerol accumulation in <i>Phaeodactylum tricornutum</i>

Supplemental Figure 1. Sequence alignment of the human PCC (HsPCC_PCC1), Ruegeria pomeroyi PCC (RpPCC_PCC1) a subunits, and the predicated PCC a subunits of Phaeodactylum tricornutum (PtPCC_PCC1) and Thalassiosira pseudonana (Tp_PCC1). Residues in the active site are indicated by arrows. Supplemental Figure 2. Sequence alignment of the human PCC (HsPCC_PCC2), Ruegeria pomeroyi PCC (RpPCC_PCC2) b subunits, and the predicated PCC b subunits of Phaeodactylum tricornutum (PtPCC_PCC2) and Thalassiosira pseudonana (Tp_PCC2). Supplemental Figure 3. Alignment of the amino acid sequences HIBCH from Homo sapiens (HsHIBCH), Arabidopsis thaliana (AtCHY1), Saccharomyces cerevisiae (ScHIBCH) and the predicted HIBCH in Chlamydomonas reinhardtii (CrHIBCH) and Phaeodactylum tricornutum (PtHIBCH). Amino acid residues identical in five, four, and three of the sequences are in red, green and blue respectively. Cleavage sites of mitochondria transit predicted by TargetP and the calculated score mTP are respectively shown by arrows and the numbers below the sequences. C-terminal peroxisome targeting signal are boxed. Asterisks above the sequence indicate proposed catalytic residues. Supplemental Figure 4. Phylogenetic relationship of the HIBCH protein family. The ML tree was generated using the MrBayes program. HIBCH are grouped into three distinct clades. Numbers above branches represent the support values (Bayesian posterior probabilities). The scale bar represents the number of nucleotide replacements per site. Supplemental Figure 5. Principal component analysis (PCA) of metabolic profiles in wild-type (WT), two HIBCH overexpression lines (hibch-OE1 and hibch-OE3) and one PCC RNAi silenced line (pcc1p). (a) Scores plot shows separation of the samples among the four groups; (b) loading plot indicated the main metabolites contributing to the separation

MOESM2 of A type-I diacylglycerol acyltransferase modulates triacylglycerol biosynthesis and fatty acid composition in the oleaginous microalga, Nannochloropsis oceanica

Additional file 2: Table S1. DGAT protein sequences used for the construction of phylogenetic tree in additional file 1: Figure S3. Table S2. Primers used in the present study. Underlined sequences designate the restriction enzyme sites. The sequences in box indicate the linker fragment introduced before GFP coding sequence

MICs of 80 nm Pd@Ag NSs against 44 cryptococcal isolates with different molecular types or from different sources compared to fluconazole and amphotericin B.

<p>MICs of 80 nm Pd@Ag NSs against 44 cryptococcal isolates with different molecular types or from different sources compared to fluconazole and amphotericin B.</p

Pd@Ag Nanosheets in Combination with Amphotericin B Exert a Potent Anti-Cryptococcal Fungicidal Effect - Fig 3

<p><b>TEM images of cryptococcal morphology before (A and B) and after (C-F) treatment with Pd@Ag NSs for 24 hours at 37°C.</b> Scale bars: 5 μm for A and C; 0.5 μm for B, D, E, and F.</p

Cell membrane permeability evaluations.

<p>Cryptococcal cells (1×10⁷ cells/mL) were cultured in the presence of calcein AM for 2 h at room temperature. After washing three times, the cells were incubated with different concentrations of Pd@Ag NSs (2–128 μg/mL) for 3 h at 37°C; AmB (2 μg/mL) and PBS were utilized as the controls. The mean fluorescence intensities (MFIs) of cellular calcein in the cells were analysed by flow cytometry. Each bar represents the mean ± SD of triplicate assays. A. control; B. Pd@Ag 2 μg/mL; C. Pd@Ag 4 μg/mL; D. Pd@Ag 8 μg/mL; E. Pd@Ag 16 μg/mL; F. Pd@Ag 32 μg/mL; G. Pd@Ag 64 μg/mL; H. Pd@Ag 128 μg/mL; I. AmB 2 μg/mL.</p

Representative large-area TEM images of Pd@Ag nanosheets with different sizes.

<p>a. 11 nm; b. 30 nm; c. 80 nm; d. 120 nm. Scale bar = 100 nm.</p

In vitro time-killing assay.

<p>The numbers of colony forming units of <i>C</i>. <i>neoformans</i> (H99) were determined after treatment with different concentrations of Pd@Ag NSs alone or combined with drugs for various periods of time. A. The Pd@Ag NSs treated group. B. The group treated with Pd@Ag NSs in combination with amphotericin B (1 μg/mL). C. The group treated with Pd@Ag NSs in combination with fluconazole (8 μg/mL). Experiments were performed in triplicate, while no drug treatment was used on the control group. The numbers in the brackets denote concentrations (μg/mL).</p

80 nm Pd@Ag NSs and amphotericin B exhibit a synergistic fungicidal effect.

<p>80 nm Pd@Ag NSs and amphotericin B exhibit a synergistic fungicidal effect.</p