Fig 1. SMA-10 is required for the intracellular trafficking of the type I receptor, SMA-6. (A) SMA-10 colocalizes preferentially with SMA-6 (type I)intracellularly: SMA-10 shows overlap with both receptors, although the fraction of colocalization with SMA-6 is higher. SMA-6::GFP and SMA-10::tagRFPwere expressed in the C. elegans intestine using the
vha-6 promoter. Young adult animals (L4+24 hrs) were imaged using the confocal microscope andcolocalization was performed. Mander's overlap coefficient for SMA-10::tagRFP and SMA-6::GFP is 0.512. (B) Loss of
sma-10 leads to intracellularaccumulation of SMA-6. SMA-6 accumulates within intracellular compartments in the
sma-10(
wk88) mutant from three regions from six animals, using atotal of 18 data points. (C) Total GFP intensity measured from worm lysates from either SMA-6::GFP or SMA-6::GFP,
sma-10(
wk88) mutant background.(D) Western blot of ubiquitination state of SMA-6 in wildtype and
sma-10 mutant background. (E) Quantification of ubiquitination data, (F) Loss of SMA-10does not affect the biosynthesis or transport of SMA-6 to the plasma membrane. Loss of
dpy-23 prevents internalization of clathrin-dependent cargos.SMA-6 biosynthesis and subsequent trafficking to the plasma membrane is unaffected in
sma-10(
wk88) animals.