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. Author manuscript; available in PMC: 2016 Oct 18.
Published in final edited form as: Nat Immunol. 2016 Apr 18;17(6):677–686. doi: 10.1038/ni.3434

Figure 3. Regulation of human and mouse autophagy gene targets by miR-33 and miR-33*.

Figure 3

(a–b) 3’UTR-luciferase reporter activity of human autophagy gene targets in HEK293 cells transfected with control (ctrl) mimics or (a) hsa-miR-33a or (b) hsa-miR-33a* (left panels). Western blotting of lysates from human THP-1 macrophages treated with control (ctrl) mimics or (a) hsa-miR-33a or (b) hsa-miR-33a* (right panels)., HSP90 is shown as an internal control (right). (c–d) 3’UTR-luciferase reporter activity of mouse autophagy gene targets in HEK293 cells transfected with control mimics or (c) mmu-miR-33 or (d) mmu-miR-33* (left panels). Western blotting of lysates from mouse peritoneal macrophages treated with control mimics or (c) mmu-miR-33 or (d) mmu-miR-33* (right panels). (e, f) mRNA levels of Atg5, Lamp1 and Prkaa1 in peritoneal macrophages transfected with (e) mmu-miR-33 or anti-miR33 or (f) mmu-miR-33* or anti-miR-33*. (g) qPCR analysis of miR-33 and miR-33* target gene mRNA expression in wild type (WT) and Mir33−/− BMDMs. (h) Quantification of miR-33 and miR-33* gene targets in GFP+ (infected) or GFP− (uninfected) alveolar macrophages isolated from mice infected with a GFP-expressing H37Rv Mtb strain for 2 weeks. *P≤0.1, **P≤0.05, ***P≤0.005 (Student’s t-test (a–d, g, h), one-way ANOVA (e, f)). Data are from one experiment representative of 3 (a–f; mean ± s.e.m) or 2 (g; mean ± s.e.m) independent experiments. Data are from 3 mice (h; mean ± s.e.m).

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