Figure 8. TRPML1 agonism prevents 1-TbAd effects on macrophages.

(A) Human M1 macrophages were pretreated with TRPML1 agonist for 1 hour, followed by incubation with 10 μM 1-TbAd for 4 hours. Scale bar: 10 μm. (B) Cells treated as in A were labeled with 10 nm immunogold and anti-LAMP1. Insets of representative images show electron-lucent compartment inclusions (upper right), while agonist-treated macrophages contained smaller electron-lucent compartments (lower right). The area (μm²) of electron-lucent lysosomal compartments was determined using the least-squares mean post test with adjustment by Tukey’s method after factorial ANOVA of a linear mixed model fit that treated each cell as a random effect variable. Scale bars: 2 μm and 500 nm (enlarged insets). (C) Human M1 macrophages loaded with the C₁₂FDG were treated as in A in biological triplicate in 2 experiments. (D) Human M1 macrophages treated as in A were subjected to BODIPY and anti-LAMP1 staining followed by BODIPY quantitation with the Kruskal-Wallis test. The results are representative of 3 experiments. Scale bar: 10 μm.