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. 2022 Jun 16;15:82. doi: 10.1186/s13045-022-01303-6

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© The Author(s) 2022

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — IR/IGF1R ratio is regulated by TRIP-Br1 in breast cancer cells. A Expression levels of TRIP-Br1, IGF1R, and IR were checked in breast normal and cancer cell lines by western blotting. β-actin was used as a loading control. B IR/IGF1R ratio was quantified using ImageJ. C, D Endogenous IR expression was assessed in MEF cells isolated from TRIP-Br1 wild-type (MEF^WT-TRIP-Br1) or knockout mice (MEF^KO-TRIP-Br1), as mentioned in the Materials and Methods (n > 3) (Additional file 3). E, F The IR protein levels from adipocytes tissue collected from TRIP-Br1 wild-type or knockout mice were evaluated by western blotting (n = 3). G, H The TRIP-Br1 and IGF1R expression levels were measured in MEF^WT-TRIP-Br1 or MEF^KO-TRIP-Br1 cells by western blotting. I, J The protein levels of TRIP-Br1 and IGF1R were checked in adipocytes tissue collected from TRIP-Br1 wild-type or knockout mice (n = 3). K The relative IR/IGF1R ratio is shown in MCF7 stable cell lines with TRIP-Br1 wild-type (MCF7^WT-TRIP-Br1) and knock-down (MCF7^KD-TRIP-Br1) cells. L The indicated protein levels were evaluated by western blotting. The expression of IGF1R and IR was co-analyzed using a co-antibody that recognizes both IGF1R and IR. M, N TRIP-Br1 or NEDD4-1 silencing RNA (siTRIP-Br1 and siNEDD4-1) were transfected into indicated cell lines and IGF1R expression was analyzed by using a western blot analysis (n > 3). O The interaction between IGF1R and TRIP-Br1 was determined by using co-immunoprecipitation assay. P, Q The representative images of NEDD4-1 and IGF1R expression were observed using a confocal microscope. The co-localization between NEDD4-1 and IGF1R was measured by counting over 50 cells in ImageJ. Data are presented as the mean ± SD (n > 50). R, S Cells were transfected with siNEDD4-1 in the absence or presence of MG132 (10 μM) for 24 h and subjected to western blotting (n = 3). The quantification results are presented as the mean ± SD (*p < 0.05; **p < 0.01; ***p < 0.005)