Supplemental Material for Bodhicharla et al., 2018
Fig. S1. Mosaic analysis of iglr-2;Ex[sur-5::gfp(NLS) pIGLR-2]worms selected for growth to adulthood on 20 mM glucose.Cell lineages and tissues carrying the extrachromosomal arrays were identified by the expression of GFP. Only worms not expressing GFP in the intestine were selected for the analysis because expression in the organ obscures expression elsewhere (Yochem et al.1998).
Fig. S2. Tissue-specific paqr-2 and fat-6 rescue and paqr-2 vitellogenin defect at L4 stage. (A)Length of 1 day old adult worms with the indicated genotypes cultivated on normal plates (NGM) or 20 mM glucose (n≥20). (B) Brood size of worms with the indicated genotypes cultivated on normal plates (n=10). (C) Photographs of worms 72 hours after placing them as L1s on normal culture plates (NGM) or culture plates containing 20 mM glucose. (D) Length of 1 day old adult worms with the indicated genotypes cultivated on normal plates (NGM) or 20 mM glucose (n≥20). (E) Tail tip phenotype of 1 day old adult worms with the indicated genotypes and cultivated on normal plates (n=100). (F) Visualization of the membranes within the developing gonad of wild type N2 and paqr-2 L4 larvae (the bright puncti in the N2 GFP image are auto-fluorescent gut granules); all L4s had normal gonads (n=50).
Fig. S3. Laurdan dye measurement of fluidity and tracking of 3H-labelled PA in transwell experiments. (A-B) Representative confocal images of HEK293 treated with PA 400 μM or vehicle for 24 h and stained with Laurdan dye. (C-D)Pseudo-color images of A and B showing GP index. Red colors indicate high membrane order, whereas blue colors indicate low order. (E)Histogram and (F)average of the GP values from HEK293 treated with PA 400 μM or vehicle for 24 h. The histogram shows how many pixels have each GP value in the region of interest (white circle, n=11 images).(G) 3H activity in the acceptor HEK293 cells exposed to different media for 24 h (H-I) Activity recorded in the transfering media after 24 h. (J) Activity registered in the donor cells at the end of the experiment. (K-L) CPM measured in the loading media after 4 h cell cultivation. (n=3 wells per condition for G-L).