Knock down of FIP1 does not impair traffic of adiponectin to the sorting endosome.

<p>Control, uninfected 3T3L1 cells, or cells stably expressing shRNAs for FIP1 were cultured and differentiated to adipocytes and used on day 10 after differentiation. Cells were fixed, permeabilized and stained with specific antibodies to detect endogenous EEA1, Rab5 and adiponectin proteins. Secondary antibodies conjugated to Alexa-350, Alexa-488 and Alexa-594 were used to visualize the proteins. Cells were imaged in a Leica DMI6000 microscope. Representative cells are shown obtained in three independent experiments. Determination of colocalization was carried out as described (Rab 5: Control (n = 9), FIP1 shRNA (n = 7); EEA1: Control (n = 10), FIP1 shRNA (n = 11) Panels d and h show the merged images.