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. 2014 Jan 20;9(1):e86373. doi: 10.1371/journal.pone.0086373

Figure 5. Quantification of localization and signaling during endocytosis.

Figure 5

Co-localization of Fgf8 is found with Clathrin, Rab7 and Lamp1, whereas co-localization of Rab5 is reduced (A). However, after stimulation of Hsc70, co-localization of Fgf8 and Rab5 is strongly increased. Three independent experiments were quantified. Cultivation of HEK293 cells after transfection with Fgf8, and Hsc70 with and without treatment of Bafilomycin A1 (B). Three independent experiments were analyzed and the ratio between activated Erk1/2 and Erk total was calculated. Cells transfected with Fgf8 showed a significant 12-fold increase in Erk1/2 phosphorylation compared to the un-transfected control. Co-transfection of Hsc70 and Fgf8 led to 8-fold increase of phosphorylation of Erk. Treatment with Bafilomycin A1 leads to a 14-fold increase of double phosphorylated Erk1/2 level. Activation by co-transfection of Fgf8 leads to a significant 19-fold increase, and co-transfection of Hsc70 and Fgf8 decreased this activation to a 10-fold activation of double phosphorylated Erk.