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. 2021 May 17;11:10416. doi: 10.1038/s41598-021-89978-8

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Levels of C3 measured by ELISA in the apical and basal conditioned media of RPE cultures showed increased levels of C3 in Efemp1^R345W/R345W:C5^+/+ and Efemp1^R345W/R345W:C5^-/- cultures compared to controls. Data from apical and basal side were combined for statistics. This represents the reality with better accuracy, since the basal conditioned media needs to be concentrated three times more, which results in more protein lost in the Amicon filter. Also C3 proteins secreted to the basal side can remain trapped inside the pores of the transwells¹⁸. Data represented as average ± SD (2-way ANOVA, *p < 0.05, **p < 0.01).