MOESM1 of Dual expression of plastidial GPAT1 and LPAT1 regulates triacylglycerol production and the fatty acid profile in Phaeodactylum tricornutum

Additional file 1: Figure S1. Protein structure of GPAT1 and LPAT1 obtained using SMART ( http://smart.embl-heidelberg.de ) and ChloroP ( http://www.cbs.dtu.dk/services/ChloroP/ ). TM, transmembrane; SP, signal peptide; LPLAT and PlsC represented the conserved domains in proteins. Figure S2. Schematic representation of the expression vectors employed in this study. GPAT1 and LPAT1 genes were cloned into the expression vectors pHY18 (A) and pHY21 (C), respectively under the control of promoter PfcpC. An omega leader sequence and “ACC” nucleotides were inserted between the promoter and the target gene for boosting protein translation. For subcellular localization, GPAT1-EGFP (B) and LPAT1-EGFP (D) were employed under the control of promoter ProPtAP. Figure S3. Subcellular localization of GPAT1 and LPAT1 in P. tricornutum cells. A, Microscopy images of a representative wild-type cell; B, Microscopy images of a representative transgenic line with co-overexpression of GPAT1 and EGFP; C, Microscopy images of a representative transgenic line with co-overexpression of LPAT1 and EGFP. From left to right, fluorescence of EGFP, autofluorescence of chloroplasts, differential interference contrast (DIC), fluorescence images overlaid on DIC image. Scale bars represent 5 μm. Figure S4. PCR validation by agarose gel electrophoresis to verify the antibiotic gene Shble (349 bp), CAT (500 bp) and endogenous gene 18s rDNA (498 bp) PCR product. V1, pHY 21 vector (containing Shble gene); V2, pHY 18 vector (containing CAT gene); OE1 and OE2, individually overexpressed transformants; WT, wild type; B, negative control; M, marker. Figure S5. Glycolipid content in overexpression lines (OE1 and OE2) and wild-type (WT) harvested at day 4 and 7. A significant difference between WT and OE lines is indicated at the p