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

. 2024 Aug 19;81(1):356. doi: 10.1007/s00018-024-05396-9

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

© 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/.

PMC Copyright notice

Fig. 4 — Tec kinase is involved in the secretion of FGF12a. a U2OS-FGF2-GFP-myc, U2OS-FGF8-GFP-myc, U2OS-FGF12a-GFP-myc and U2OS-FGF1-myc cells were serum-starved at 37 °C for 24 h. The media were exchanged, and the cells were incubated at 42 °C for 2 h. The media from above the cells were collected, centrifuged, and incubated with anti-myc tag magnetic beads. The eluted proteins and lysates were analyzed by SDS-PAGE and western blotting using anti-GFP antibody. The graph shows the relative secretion of proteins quantified using densitometric measurements in Fiji software. Student’s t-test was applied for statistical analysis; ns p > 0.05, ** p ≤ 0.01, *** p ≤ 0.001. b U2OS-FGF2-GFP-myc, U2OS-FGF8-GFP-myc and U2OS-FGF12a-GFP-myc cells were transfected with siRNA targeting Tec kinase or scramble siRNA (control) and, after 48 h, the cells were serum-starved at 37 °C for 24 h. After the media exchange, the cells were incubated at 42 °C for 2 h. The media from above the cells were collected, centrifuged and incubated with anti-myc tag magnetic beads. The eluted proteins and lysates were analyzed by SDS-PAGE and western blotting using anti-GFP antibody. The anti-Tec antibody was used to verify knock-down effectiveness. The graph shows the relative protein secretion quantified using densitometric measurements in Fiji software. Student’s t-test was applied for statistical analysis; ns p > 0.05, ** p ≤ 0.01. c Sequence alignment of wild type FGF12a and designed mutant variants. d U2OS cells were transiently transfected with plasmids encoding FGF12a-GFP-myc, FGF12aY126A-GFP-myc, FGF12aH1-GFP-myc, FGF12aH2-GFP-myc, FGF2-GFP-myc and FGF2Y82A-GFP-myc. 48 h after transfection, the media were exchanged and the cells were serum-starved at 37 °C for 24 h. The media were exchanged again and cells were incubated at 42 °C for 2 h. The media from above the cells were collected, centrifuged and incubated with anti-myc tag magnetic beads. The eluted proteins and lysates were analyzed by SDS-PAGE and western blotting using anti-GFP antibody. Graphs show the relative protein secretion quantified using densitometric measurements in Fiji software. Student’s t-test was applied for statistical analysis; **p ≤ 0.01, ***p ≤ 0.001