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

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.