Fig. 1

Overexpression of FAX1/FAX2 and ABCA2 in Chlamydomonas. a Schematic representation of the transformation vectors for generation of FAX1/FAX2 (left panel), and FAX1/FAX2/ABCA2 (right panel) overexpressing strains. Pro, Hsp70A/RbcS2 promoter; 2A, FMDV 2A peptide; Ter, Chlamydomonas RbcS2 terminator. b, c Reverse-transcription PCR (RT-PCR) analysis of the expression of FAX1/FAX2, and ABCA2 in the transformants. PL, parental line (FAX1-FAX2-35). Housekeeping gene used is RACK1 (Cre06.g278222, receptor of activated protein kinase C 1 initially described as CBLP). The PCR products amplified with the primers (FAX1-F/FAX2-R and ABCA2-F1/ABCA2-R1) at 28 cycles were shown, respectively. This is a representative of three independent experiments. d Quantitative RT-PCR analysis of the expression level of ABCA2 in FAX1/FAX2/ABCA2 transformant. The qRT-PCR results were normalized by the level of RACK1 expression. PL, parental line (FAX1-FAX2-35). Error bars represent standard errors based on three biological replicates (independent cultures) with three technical replicates each. Asterisks indicate statistically significant changes compared to the parental line FAX1-FAX2-35 by paired-sample Student’s t test (**P ≤ 0.01)