Related Experiment Video
Updated: May 26, 2026
Polarization of M1 and M2 Human Monocyte-Derived Cells and Analysis with Flow Cytometry upon Mycobacterium tuberculosis Infection
Published on: September 18, 2020
[Effects of Mycobacterium tuberculosis Hsp16.3 protein on the autophagy function of mice macrophages]
Chang-hong Shi1, Ying Jiang, Yong Zhao
1Laboratory Animals Center, Fourth Military Medical University, Xi'an 710032, China.
Aim:
To investisate the inhibition of Hsp-16.3 on the autophagosomes formation of macrophages.
Methods:
Mouse RAW264.7 macrophages were induced by rapamycin (50 ng/μL) following infection with M.tuberculosis H37Rv strains, thereafter, co-incubated with Hsp16.3 protein (25 μg/mL). The effects of Hsp16.3 protein on the autophagosomes formation was observed with transmission electron microscope. The expression of autophagy-related genes (atg8) for macrophages was detected by Western blotting.
Results:
It was found that rapamycin-induced autophagy of macrophages infected with M.tuberculosis H37Rv enhanced localization of mycobacteria with autophagosomes. Hsp16.3 protein inhibits autophagosome formation and affects M.tuberculosis survival inside infected macrophages. Furthermore, Hsp16.3 protein significantly increased M.tuberculosis colony forming units (CFU), and decreased the expression of microtubule-associated protein light chain-3 (LC3) expression level (P<0.05).
Conclusion:
The results showed that Hsp16.3 protein inhibits the formation of autophagosomes by regulating the expression of LC3 protein.
