Fig 5. Sterol biosynthesis from glucose, threonine and acetate requires SCP2-thiolase, and fatty acid biosynthesis from acetate requires ASCT.

(A) Western blot analyses of the parental (WT), knock-out and tetracycline-induced (.i) or non-induced (.ni) mutant cell lines with the immune sera indicated in the right margin. (B) The top and lower parts represent the relative incorporation of radio-labelled carbon sources (D-[U-¹⁴C]-glucose, L-[U-¹⁴C]-threonine, [1-¹⁴C]-acetate and L-[U-¹⁴C]-leucine) into fatty acids and sterols, respectively, of tetracycline-induced (.i) and non-induced (.ni) ^RNAiSCP2, Δach and Δasct cell lines, compared to the parental cell line (PCL). [¹⁴C]-labelled fatty acid methyl esters and sterols were separated by HPTLC and analyzed as described in the Materials and methods section. Data are normalized with the parental cell (PCL) values with an arbitrary value of 100 for the PCL samples, which is represented by a horizontal dashed lane. Error bars indicate mean ± SD of 3 biological replicates. nd: not detectable.