Fig. 5

BMP inhibitors together with Ym155 or phenformin induce AMPK/LKB1 dependent nuclear localization of AIF. A, B AIF immunostaining of H1299 and A549 cells treated with JL5 and Ym155 alone and in combination for 24 h. C, D AIF immunostaining of H1299 and A549 cells treated with JL189 and Ym155 alone and in combination for 24 h. E, F AIF immunostaining of H1299 and A549 cells treated with JL5 and phenformin alone and in combination for 24 h. Representative immunofluorescent images of AIF (green) with nucleus stained with DAPI (blue). Arrow depicts cell with nuclear staining for AIF. G AIF immunostaining of H1299 cells transfected with siRNA control or siRNA BMPR2 then treated with DMSO or Ym155 20 nM for 24 h. H AIF immunostaining of primary lung carcinomas treated with JL5 and Ym155 alone and in combination. Graphs depict representative experiments of each condition, demonstrating percentage of cells with AIF localized to the nucleus. Approximately 50–70 cells from random high-power fields were counted. I Western blot of H1299 transfected with siRNA scrambled control and siRNA for LKB1. J Localization of nuclear localization of AIF of treated H1299 cells following transfection with siRNA scrambled control and siRNA for LKB1. Data represents mean of 3 experiments. K Nuclear localization of AIF in A549-Puro and A549-LKB1 treated for 54 h. Graphs represent the mean of 2 experiments with approximately 100 cells or more counted per condition. Scale bars are equal to 10 μm