Future investigations could focus on genic markers and examine how their characteristics are associated with measures of genetic variation.
In this study, we focused on EpCAM, CD24, CD44, CD133, CXCR4 and ALDH1 as representative pancreatic CSC markers and examined the effect of NACRT on pancreatic CSCs.
To define the nature of the dispersed GM130-positive structures in parietal cells more precisely, we employed additional Golgi markers and examined the relationship between cis- and trans-cisternae in the Golgi fragments.
To improve filtering, we will also genotype the EPGP full siblings for a linkage panel of markers and examine whether regions containing potentially pathogenic variants in the sequenced sibling are also inherited in the other sibling.
In any case, the geographic congruence between K = 4 AFLP clusters and the mtDNA phylogenetic tree provides a unique opportunity to compare patterns of divergence between the two types of markers and examine the behaviour of each one at Z. vivipara contact zones in the region.
To examine this issue, we transformed lsm12Δ and pbp4Δ strains with a plasmid expressing Pab1-GFP (stress granule marker) and Edc3-mCherry (P-body marker), and examined the formation of stress granules and P-bodies during glucose deprivation.
The samples were washed three times with PBS, incubated for 5 min with 1.3 nM Hoechst 33342 (DNA marker), and examined under a Leica SP5 confocal laser microscope (Leica Microsystems, Wetzlar, Germany).
PCR reaction products were then separated on 4% agarose gels containing ethidium bromide with 100 bp DNA Ladder Markers (Bioline) and examined under ultraviolet illumination to identify the distinct bands.
To study the subcellular localization of MANF and its mutants, we coimmunostained the microinjected healthy NGF-maintained SCG neurons with antibodies to MANF and to either the ER markers protein disulfide isomerase (PDI) or 78 kDa glucose-regulated protein (GRP78), or to the Golgi marker GM130, and examined the neurons using confocal microscopy.
To demonstrate that vacuolar PIN2-eGFP was derived from the PM via endocytic vesicle trafficking, we pulse-labeled 5-day-old light-grown PIN2-eGFP seedlings with an endocytic marker, FM4-64 and examined the kinetics of FM4-64 endocytosis and PIN2-eGFP vacuolar accumulation in a time course after shifting seedlings to dark.
