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

. 2023 Nov 17;14:7476. doi: 10.1038/s41467-023-43292-1

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

© The Author(s) 2023

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. 8 — a Quantification by qPCR of the relative p21 mRNA in the GDF11^KO and WT Neuro-2a cells (n = 3 clones). b–e Immunofluorescence representative images (b) and quantification of the number of p21⁺ cells per field (c, n = 5 fields/group), the proportion of p21⁺ cells (d, n = 6 fields/group) or the average gray value of p21 per cell (e, GDF11^KO, n = 420 cells; WT, n = 280 cells) in the GDF11^KO and WT Neuro-2a cells. Scale bar, 25 μm. Examples of the p21⁺ cells are indicated by double arrowheads. f–h The same snRNA-seq data were used, as described in Fig. 5a. f Rank for regulons in the EN based on regulon specificity score (RSS). The EN were obtained from the Cg2 of the “KO” mice and the “Ctrl” mice aged 4–5 M. g Regulons activity analysis based on area under the curve (AUC) in the identified cell types in snRNA-seq of the “KO” mice and the “Ctrl” mice aged 4–5 M. The activity of regulon Smad3 (highlighted in red) is high in the EN. h Cytoscape network visualization of genes including GDF11, Cdkn1a (p21), Smad2, Smad3 (highlighted in red) and their transcription factors (TFs, yellow). i–m Representative images (i and l) and quantification by densitometry of western blot analysis of Smad2 (j), phosphorylated Smad2 (pSmad2, k) and Smad3 (m) in the total protein extracted from the GDF11^KO and WT Neuro-2a cells (n = 3 biological repeats/group). n ChIP-qPCR assessment of the enrichment of Smad2 at the promoter of Cdkn1a/p21 in the GDF11^KO and WT Neuro-2a cells (n = 3 biological repeats/group). o A proposed working model for loss of GDF11 on cellular senescence. Loss of GDF11 upregulates pSmad2, enhances nuclear entry of Smad2/3 tricomplex and then Smad2 binds to the promoter of p21 and promotes the pro-senescence factor p21 transcription, and eventually causes cellular senescence. Data are presented as mean ± SEM. *P < 0.05, **P < 0.01 and “ns” indicates not significant. a (P = 0.0037), c (P = 0.0033), d (P = 0.0157), e (P < 0.0001), j (P = 0.6648), k (P = 0.0040) and m (P = 0.0299), unpaired two-tailed t test. n (IgG: WT versus GDF11^KO, P = 0.57; Smad2: WT versus GDF11^KO, P < 0.001), two-way ANOVA with Sidak’s test. Source data are provided with this paper.