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. 2024 May 3;43:133. doi: 10.1186/s13046-024-03047-2

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© The Author(s) 2024

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/. 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. 7 — FCN3 inhibits IR activation by promoting the dephosphorylation of IR-β and suppressing pro-IR cleavage

(A) Immunoblots of p-IR-β and IR-β in nsFCN3-overexpressed MHCC97-H and YY-8103 cells. GAPDH was used as a loading control. (B-C) Quantitative analysis of p-IR-β/IR-β ratio in nsFCN3-overexpressed MHCC97-H (B) and YY-8103 (C) cells. (D-E) Immunoblots (D) and quantification (E) of p-IR-β and IR-β in FCN3-overexpressed YY-8103 cells. GAPDH was used as a loading control. (F-G) Immunoblots of FCN3 and HA-tagged IR after immunoprecipitation of HA-tagged IR from HEK-293T cells that transfected with FCN3 (F) or nsFCN3 (G) and IR-HA. (H-I) Immunoblots of IR-β, PTP1B and FCN3 after immunoprecipitation of Flag-tagged FCN3 from HEK-293T cells that transfected with FCN3-Flag (H) or nsFCN3-Flag (I). (J) Immunoblots of PTP1B, FCN3 and HA-tagged IR after immunoprecipitation of HA-tagged IR from indicated HEK-293T cells. (K) mRNA level of IR in nsFCN3-overexpressed YY-8103 cells. (L-M) Immunoblots (L) and quantification (M) of pro-IR and IR-β in nsFCN3-overexpressed YY-8103 cells. GAPDH was used as a loading control. (N-O) Immunoblots of pro-IR and FCN3 after immunoprecipitation of Flag-tagged FCN3 from HEK-293T cells that transfected with FCN3-Flag (N) or nsFCN3-Flag (O). (P) Immunoblots of Furin, FCN3 and HA-tagged pro-IR after immunoprecipitation of HA-tagged IR from indicated HEK-293T cells. (Q) Immunoblots of indicated proteins in HEK-293T cells. GAPDH was used as a loading control

Data are from one representative experiment of three independent experiments (A-E and K-M). Data are presented as mean ± SD. Significance was assessed by Student’s t test (B, C, E, K, M). **p < 0.01, ***p < 0.001 compared with the control group. ns, not significant