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. 2022 Mar 31;12:730530. doi: 10.3389/fonc.2022.730530

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Copyright © 2022 Chen, Wang, Liu, Liang, Wang, Wu, Xu, Liang, Ma and Gao

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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N-glycosylation increases stability of TIM-4. (A) Western Blot was used to detect the stability of TIM-4 before and after the removal of N-glycosylation. (B) Western Blot was used to detect the degradation pathway of TIM-4 before and after the removal of N-glycosylation. (C) Ubi-HA was transfected into A549-LV-TIM-4-Flag cells, and TM or MG132 was used to treat cells. TIM-4 was precipitated by anti-Flag, and Western Blot was used to detect the ubiquitination of TIM-4 before and after the removal of N-glycosylation. (D) A549 cells were transfected with pcDNA3, TIM-4-HA or N291Q-HA respectively and treated with MG132. TIM-4 was precipitated by anti-HA, and Western Blot was performed to detect the ubiquitination of TIM-4. **p < 0.01, ***p < 0.001, by Student’s t-test.