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. 2021 Dec 20;13(1):11. doi: 10.1038/s41419-021-04458-1

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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 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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Fig. 6 — A Detection of phosphorylated Smad2 (pSmad2) and total Smad2 (Smad2) in cell lysates from wild-type and tmem165-knockout mouse ATDC5 cells (n = 3). B Fold changes of serpine expression in tmem165-knockout cells normalized to wild-type mouse ATDC5 cells. C Fold changes of TGF-β reporter activity in tmem165-knockout cells normalized to wild-type mouse ATDC5. D Detection of phosphorylated Smad2 (pSmad2) and total Smad2 in cell lysates from control and TMEM165-deficient CDG patient fibroblast cells (n = 3). E Fold changes of serpine expression in TMEM165-deficient fibroblasts normalized to normal fibroblast cells. F Detection of p-Smad2 in cell lysates from wild-type and tmem165-knockout mouse ATDC5 cells and (G) from normal and TMEM165-deficient CDG patient fibroblast cells treated or not with TGFβ1 (1 ng/ml) for 1 hour (n = 3). H Fold changes of TGFβR1 and TGFβR2 expression in tmem165-knockout mouse ATDC5 cells normalized to wild-type mouse ATDC5 cells. I Detection of TGFβR2 in cell lysates from wild-type and tmem165-mutant mouse ATDC5 (n = 3). J Fold changes of TGFβR1 and TGFβR2 expression in TMEM165-deficient fibroblasts from CDG patient normalized to normal fibroblast cells. K Detection of TGFβR2 in cell lysates from normal fibroblasts and TMEM165-deficient CDG patient fibroblast cells (n = 3). L Fold changes of asporin expression in tmem165-knockout cells normalized to wild-type mouse ATDC5 cells and in TMEM165-deficient fibroblast cells normalized to normal fibroblast cells. M Detection of asporin in conditioned medium of wild-type and tmem165-knockout mouse ATDC5 cells and of normal fibroblasts and TMEM165-deficient CDG patient fibrobast cells. β-actin was used as loading control (n = 3). qPCR values were normalized for the housekeeping gene ribosomal protein S29 and are expressed as the relative expression compared with control. Data are expressed as mean ± S.D. Statistical analysis was performed with an unpaired Student’s t test (n = 3; *p < 0.05; **p < 0.01). Representative images from three independent experiments are shown.