Fig 1: TRAPP II complex integrity is affected by TRAPPC10 variants and in TRAPPC10-/- cells.Yeast cells transformed with pGBKT7 or pGBKT7 expressing either wild type TRAPPC10, the p.Gly1131Valfs*19 variant or the p.Pro929Leu variant (A), or pGADT7 or pGADT7 expressing either wild type TRAPPC10, the p.Gly1131Valfs*19 variant or the p.Pro929Leu variant (B) were mated with yeast expressing either pGBKT7-TRAPPC2L (A) or pGADT7-TRAPPC2L (B). Serial dilutions of the diploid cells were grown on SC medium lacking leucine and tryptophan (DDO) or SC medium lacking leucine, tryptophan, histidine and adenine (QDO). (C) Diploid cells from (A) and (B) were quantitatively tested for ß-galactosidase activity. Units of activity were calculated according to the following formula: (OD420 x 1000)/(OD600 x time in hours). Note that western analysis revealed that all forms of TRAPPC10 and the two variants were expressed in the yeast cells. (D) Lymphoblastoid cells from control or an individual homozygous for the TRAPPC10 p.(Gly1131Valfs*19) variant were lysed and probed for the TRAPP proteins indicated and for tubulin as a representative loading control. The lysates were fractionated on a Superose 6 size exclusion column. Fractions of 0.5ml were collected. Neighbouring fractions were pooled and fractionated by SDS-PAGE and probed for the indicated TRAPP proteins. (E) Wild type HEK293 or TRAPPC10-/- cells were lysed and probed for the TRAPP proteins indicated and for tubulin as a representative loading control. The lysates were fractionated on a Superose 6 size exclusion column. Fractions of 0.5ml were collected. Neighbouring fractions were pooled and fractionated by SDS-PAGE and probed for the indicated TRAPP proteins. The fractionation of molecular size standards are indicated above the top-most panel of the size exclusion portion of (D) and (E).