Figure S2.

Polyisoprenoid LCMS chromatograms and changes in DHRSX- and SRD5A3-deficient fibroblasts, related to Figure 2

(A) Representative mass spectra (right) and extracted ion chromatograms (left) for the indicated m/z, obtained from standards for polyprenal (PAL), polyprenol (POL), dolichal (DAL), dolichol (DOL) and a mixture of all four species (bottom).

(B) Representative extracted ion chromatograms of indicated m/z values in WT, DHRSX KO and SRD5A3 KO HAP1 cells showing accumulation of polyprenal (PAL) and polyprenoic acid (POL-COOH) only in SRD5A3 KO cells and polyprenol (POL) in both DHRSX and SRD5A3 KO cells. Dolichol (DOL) levels are reduced in both DHRSX and SRD5A3 KO cells. The [M+2] peak is shown to reduce confounding effects of increased polyprenal M+4 levels on signals for dolichol.

(C) Polyisoprenoids in dermal fibroblasts collected from DHRSX-CDG patients (P1 & P3), compared to those from three controls. Data is the total ion count (TIC)-normalized area under the curve (AUC) (means ± SEM; n = 4). § p < 0.05 compared to every control; # p < 0.05 compared to one of the controls. Only the species with 19 isoprenyl units are shown for clarity. See Table S2 for all isoprenyl chain lengths. Note the up to 100-fold difference in scale between panels.

(D) Polyisoprenoids in dermal fibroblasts collected from six SRD5A3-deficient individuals (SRD5A3-P1-6), compared to those from three controls. Data is TIC-normalized AUC (means ± SEM; n = 4). § p < 0.05 compared to every control; # p < 0.05 compared to one of the controls. Only the isoprenoid species with 19 isoprenyl units are shown for clarity. See Table S2 for all isoprenyl chain lengths.