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. 2018 Jan 22;8:1368. doi: 10.1038/s41598-018-19542-4

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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EGF-induced EGFR signaling and cell growth are impaired in GGA2-depleted cells. (a) Control (siCtrl) and GGA2-depleted (siGGA2-#1 and siGGA2-#2) ARPE-19 cells were treated with (+) or without (−) 10 nM EGF for 5 min and were then lysed for western blot analyses with antibodies against EGFR, phosphorylated MAPK (P-MAPK), MAPK, GGA2, or tubulin. Uncropped blots are presented in Supplementary Fig. 7. (b) Increase in the ratio of P-MAPK to MAPK after EGF stimulation was calculated and normalized to the value of siCtrl. Mean (±SD) of three experiments were plotted. Statistical difference between the control and each GGA2 siRNA was identified using Student’s t-test (**P < 0.01 and ***P < 0.001). (c) Control (Ctrl) or GGA2-depleted (shGGA2) LoVo cells were transplanted into nude mice. Tumors were excised at 4 weeks after transplantation. (d) Tumor weights and volumes were measured at the indicated time points and displayed as dot plots (mean [horizontal bar] ± SD). Statistical analyses were performed using Mann-Whitney U-test; ***P < 0.001.