MDMs from LTBI^– healthy donors were transfected with siRNA targeting CES-1, Siglec-14, RPS-26, RGCC1, and ANXA1 and control siRNA. The siRNA-transfected MDMs were infected with H37Rv at an MOI of 2.5. (A) After 5 days, the supernatant was aspirated, and the MDMs were lysed. The supernatant was centrifuged to pellet the bacteria, and the pellets were added to the cell lysates. The bacterial suspensions were ultrasonically dispersed, serially diluted, and plated in triplicate on 7H10 agar. The number of resultant colonies was counted after 3 weeks. The P values were determined by unpaired 2-tailed t test. The mean ± SD is shown. The means and SDs are shown for the number of CFUs per well. (B) The number of apoptotic MDMs was determined by flow cytometry. (C) The percentage of LC3⁺ MDMs was determined by flow cytometry. (D) Freshly prepared MDMs were infected with converter or nonconverter plasma opsonized or unopsonized M. tuberculosis H37Rv at an MOI of 2.5. The P values were determined by 1-way ANOVA with Tukey’s multiple comparisons test. Means, SDs, and P values are shown. (E) Control or Siglec-14 siRNA–transfected MDMs were infected with H37Rv at an MOI of 2.5. After 72 hours, RNA was isolated from MDMs, and a PCR array was performed for antimicrobial peptides. Data were normalized (z score) and centered using the Clustvis program. (F) CD14⁺Siglec-14⁺ and CD14⁺Siglec-14^– cells were magnetically sorted from the PBMCs of healthy donors (n = 6). Sorted cells were infected with M. tuberculosis H37Rv at an MOI of 2.5. At 2 hours and 5 days postinfection, the number of bacterial colonies was determined as outlined above. (G) Freshly isolated PBMCs from LTBI^– healthy donors (n = 3) were cultured in the presence or absence of γ-M. tuberculosis. After 72 hours, CD14⁺Siglec-14⁺ and CD14⁺Siglec-14^– cells were sorted through magnetic labeling, and RNA was isolated. Quantitative real-time PCR was performed to determine the mRNA expression level of antimicrobial peptides. The P values were determined by 1-way ANOVA with Tukey’s multiple comparisons test. Mean values, SDs, and P values are shown.