Figure 4.

miR-25 directly targets NPC1

(A) Predicted binding sites of miR-25 and NPC1 3′UTR.

(B) Luciferase activity of HEK293T cells transfected with either PmirGLO-NPC1-WT-UTR or PmirGLO-NPC1-MUT-UTR plus control mimic and miR-25 mimic.

(C) THP-1 cells pretreated with control mimic, miR-25 mimic, control inhibitor or miR-25 inhibitor were infected with Bacillus Calmette–Guérin for a specified period of time, and qRT-PCR detection of NPC1 levels was performed,

(D and E) western blotting of the lysate was performed to detect the amount of NPC1. (F and G) THP-1 cells pre-transfected with control siRNA or NPC1 siRNA were infected with BCG or H37Rv for a specified period of time, and then CFU detection was performed, indirect immunofluorescence (IF) analysis of the co-localization of autophagosomes (LC3, green) and lysosomes (LAMP-1, red) bar, 10 μM (H).

(I) Pearson correlation coefficients (PCCs) of images of internalized Alexa Fluor 488-LC3 and Alexa Fluor 594-LAMP1 in THP-1 cells. Data are presented as means ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.005, ns, not significant.