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. 2014 Jun 17;79(1):139–149. doi: 10.1111/tpj.12547

Figure 2.

Figure 2

Influence of boron on glycosylinositol phosphorylceramide (GIPC) extraction. (a) A cloudy layer was observed during butanol/water phase-partitioning of a GIPC-enriched lipid sample extracted with neutral ethanol from Rosa cell cultures that had been grown in the usual B concentration (i, iv). This cloudy layer disappeared in the presence of 0.1 m HCl (iii, vii), 10 mm βMCD (ii), or 6 mm borate buffer, pH 9.2 (vi). The horizontal arrow indicates the slight cloudy layer left in the presence of βMCD (butanol above). In contrast, 6 mm ammonium buffer, pH 9.2 (v), only led to a partial disappearance. (b) TLC of the different phases after butanol/water phase-partitioning of a GIPC-rich lipid extract from Rosa cell cultures grown in media with (B+) or without boron (B−). The lipids had been extracted in 70% ethanol that contained 0.1 m HCl (H+) or lacking acid (H−). BP, butanol phase; CL, cloudy layer; AP, aqueous phase; Suc, sucrose (marker). In lanes 9 and 10, 10 mm βMCD was present during the partitioning step. Lipids labelled on lane 10: bands 1–3, as in Figure1; band 4, (Pent)2-(Hex)2-HexA-Ins-P-Cer.