Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2019 Jun 20;2:227. doi: 10.1038/s42003-019-0478-3

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2019

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/.

PMC Copyright notice

Fig. 4 — MCRIP1 promotes the expression of SP-B and SP-C by disrupting the CtBP-Foxp interactions. a, b MCRIP1 inhibits Foxp-CtBP interactions. U2OS cells were transiently transfected as indicated. a Flag-CtBP1 was immunoprecipitated, and co-precipitated Myc-MCRIP1 and Myc-Foxp2 were probed with an anti-Myc antibody. Increasing amounts of Myc-MCRIP1 expression vector were used. b Myc-CtBP2 was immunoprecipitated, and co-precipitated Flag-MCRIP1, Flag-Foxp1, or Flag-Foxp2 were probed with an anti-Flag antibody. c Co-precipitation of endogenous CtBP1 and Foxp1 in lung lysates from Mcrip1^+/+ and Mcrip1^−/− mice. Endogenous Foxp1 was immunoprecipitated and co-precipitated CtBP1 was detected by immunoblotting. a–c The levels of protein expression are shown in the lower rows. d, e MCRIP1 inhibits the repression of the SP-B promoter induced by Foxp2 d or CtBP2. e A549 cells were transfected with the SP-B-luciferase reporter plasmid (SP-B Luc) and the pMCRIP1 plasmid, together with pFoxp2 d or pCtBP2 e as indicated, and SP-B promoter activity was analyzed (fold induction). f SP-B and SP-C promoter assays. A549 cells were transfected with siRNA targeting human MCRIP1, cultured for 48 h, and then re-transfected with SP-B-Luc or SP-C-luciferase reporter plasmid (SP-CLuc). After an additional 48-h incubation, SP-B, or SP-C promoter activities were analyzed. d–f Data are means ± SEM from three independent experiments performed in triplicate. g MCRIP1 siRNA-mediated depletion decreases the mRNA expression of SP-B and SP-C. MLE12 cells were transfected with siRNAs targeting mouse Mcrip1 (#1 or #2). Total RNA was then extracted and analyzed for endogenous SP-B (left) or SP-C (right) mRNA expression (fold change) using qRT-PCR. h, i Mcrip1-KO MLE12 cells were established using the CRISPR/Cas9 approach with two different guide RNAs (KO#1 or KO#2). h The expression levels of endogenous MCRIP1 and SP-B proteins were analyzed by immunoblotting. β-actin, loading control. i The expression levels of SP-B and SP-C mRNAs were analyzed using qRT-PCR. g, i The data represent the mean ± SEM from three independent experiments