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. 2021 Feb 24;6(1):e00036-21. doi: 10.1128/mSphere.00036-21

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Copyright © 2021 Madhavan et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

PMC Copyright notice

FIG 4 — DP and CI994 induce autophagy in human macrophages. (A and B) DP and CI994 treatment of infected macrophages upregulated the expression of the autophagy-related genes BECN1 and ATG5. Differentiated human THP-1 cells were infected with M. tuberculosis H37Rv for 4 h and treated with DP and CI994 or rapamycin (Rapa). The mRNA levels of autophagy-related genes ATG5 (A) and BECN1 (B) (normalized to β-actin expression) in THP-1-derived macrophages were measured by qPCR. Results are shown from 3 independent experiments (mean ± SD). *#, expression of BECN1 and ATG5 is significantly different from that in uninfected (UIF) and infected (IF) samples, respectively (P ≤ 0.05). (C) Western blotting of ATG5, Beclin 1, and β-actin. (D) Densitometric analysis of ATG5 and Beclin 1 bands normalized with that of β-actin. Each value represents mean ± SD from triplicate measurements. *#, band intensity of ATG5 and Beclin 1 is significantly different from that in uninfected (UIF) and infected (IF) samples, respectively (P ≤ 0.05). (E) Immuno-cytochemical imaging shows levels of Beclin 1 in variously infected macrophages at 24 hpi.