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. 2016 Nov 1;7(48):79203–79216. doi: 10.18632/oncotarget.13008

Figure 2. In absence of ligand stimulation, increased levels of EGFR in EHD3-expressing cells are associated with decreased ubiquitination and less recruitment of the protein Cbl.

Figure 2

(A) Protein extracts from U251tetEhd3 cells induced (+) or not (−) with Dox for 1–3 days, were used to immunoprecipitate (IP) ubiquitinated proteins and subsequently assessed by immunoblotting (IB) using an anti-EGFR antibody. A longer exposure is shown for a better appreciation of the effects at day 3. The IgG IP control was performed with samples collected on day 2. (B) The histogram depicts densitometric quantitation of a representative immunoblot and the results are shown as relative units (r.u.) of the EGFR/GAPDH ratio in Dox-induced versus the control without Dox treatment, the latter being at a 100% value. C) An EGFR IP was performed on similar samples as in A, and subject to immunoblotting to assess levels of EGFR, Cbl and Shc.