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

. 2020 Jul 1;9(7):1591. doi: 10.3390/cells9071591

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

© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

ATRA mediates metabolic changes in LPS-primed MΦs. The cells were pre-treated with or without ATRA, then primed with LPS for 6 h, and subsequently subjected to the mitochondria stress test using a Seahorse XF96 Analyzer. (A) Real-time kinetics measurement of the oxygen consumption rate (OCR) during sequential treatment with oligomycin (Oligo), carbonylcyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP), and antimycin A + rotenone (A+R). Representative results are shown. Bar graphs represent the calculated basal and maximal OCR, and ATP-coupled respiration and spare respiratory capacity. (B) Real-time kinetics measurement of the extracellular acidification rate (ECAR) after sequential treatment of glucose (Glu), oligomycin (Oligo), and 2-deoxyglucose (2-DG). Representative results are shown. Bar graphs represent calculated ECAR and glycolytic capacity obtained from the glycolytic stress test. Wave Desktop software was used for data analysis. (C) Relative gene expression of HK2 was measured by qPCR. The expression was normalized to the reference gene (human cyclophilin) expression. (D) MΦs were pretreated with 3-bromopyruvate (3BP) (80 µM) 1 h before LPS priming and subsequently incubated with ATP (5 mM) for 45 min., and then the cell culture supernatants were collected, and the secretion of IL-1β was assessed by ELISA. C, control (mock-treated cells). The data was obtained from at least four healthy donors. All results are shown as means ± SEM. (* p < 0.05, ** p < 0.01, *** p < 0.001). +, −, presence or absence of indicated substance, respectively

ATRA mediates metabolic changes in LPS-primed MΦs. The cells were pre-treated with or without ATRA, then primed with LPS for 6 h, and subsequently subjected to the mitochondria stress test using a Seahorse XF96 Analyzer. (A) Real-time kinetics measurement of the oxygen consumption rate (OCR) during sequential treatment with oligomycin (Oligo), carbonylcyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP), and antimycin A + rotenone (A+R). Representative results are shown. Bar graphs represent the calculated basal and maximal OCR, and ATP-coupled respiration and spare respiratory capacity. (B) Real-time kinetics measurement of the extracellular acidification rate (ECAR) after sequential treatment of glucose (Glu), oligomycin (Oligo), and 2-deoxyglucose (2-DG). Representative results are shown. Bar graphs represent calculated ECAR and glycolytic capacity obtained from the glycolytic stress test. Wave Desktop software was used for data analysis. (C) Relative gene expression of HK2 was measured by qPCR. The expression was normalized to the reference gene (human cyclophilin) expression. (D) MΦs were pretreated with 3-bromopyruvate (3BP) (80 µM) 1 h before LPS priming and subsequently incubated with ATP (5 mM) for 45 min., and then the cell culture supernatants were collected, and the secretion of IL-1β was assessed by ELISA. C, control (mock-treated cells). The data was obtained from at least four healthy donors. All results are shown as means ± SEM. (* p < 0.05, ** p < 0.01, *** p < 0.001). +, −, presence or absence of indicated substance, respectively