Fig. 5

JL5 induces cytosolic trapping of BMPR2 in lung cancer cells and C. elegans. a H1299 cells were treated with DMSO, DMH1 2.0 μM, or JL5 2.5 μM for 24 h and immunofluorescent imaging performed for BMPR2 (green) and nuclear staining with DAPI. Only cells treated with JL5 demonstrated BMPR2 trapped within cytoplasmic vesicles. Studies were performed at least 3 times with the same results. b Various concentrations JL5 were applied to L1 C. elegans bearing a germline-integrated spp-9::GFP transgene. After 72 h animals were imaged at 5x magnification on a standard epifluorescent microscope. Imaging quantification was performed using the open-source Fiji Software. Graphs represent population spread with mean +/− SEM. A minimum of 60 animals were quantified for each condition. A one-way ANOVA was performed to compare differences in mean intensity across conditions. JL5 increased GFP fluorescent intensity indicating a decrease in BMP signaling. c JL5 traps BMPR2 (daf-4) within an intracellular compartment. C. elegans bearing a daf-4::GFP (BMPR2) transgene were treated with DMSO or JL5 for 72 h and live animals examined by confocal microscopy. Arrows demonstrate abnormal daf-4::GFP (BMPR2) accumulation within vesicles close to the basolateral membrane. Sixty live animals were examined for each condition, which were performed twice