Fig. 4. Increased levels of PI3P in C. elegans caused by knockdown of ceMTM3 expression by RNAi.

PI3P was extracted from N2 worms cultured on NGM plates containing E. coli cells carrying vector control or ceMTM3bF1 and then fractionated on thin layer chromatography plates as described in Methods and Materials. Equivalent amounts of samples together with standard PI3P were spotted on a nitrocellulose membrane. The membrane was incubated with a GST-ceMTM3CT followed by detection with anti-GST antibody and HRP-conjugated secondary antibody. Capture of immunoblot images and quantification of band signals were carried out by using FluorChem SP imaging system from Alpha Innotech. Error bars denote standard deviation (n ≥ 4).