Skip to main content
. Author manuscript; available in PMC: 2020 Aug 31.
Published in final edited form as: Cell Rep. 2019 Mar 26;26(13):3613–3628.e6. doi: 10.1016/j.celrep.2019.02.102

Figure 4: FLCN depletion in macrophages enhances their energy metabolism and phagocytic potential.

Figure 4:

(A) Glucose production and (B) lactate consumption levels measured using NOVA Bioanalysis flux analyzer in empty vector (EV) or shFLCN RAW264.7 at basal level. (C, D) Extracellular acidification rate (ECAR) and (E, F) oxygen consumption rate (OCR) of EV or shFLCN RAW264.7 at basal level as measured by Seahorse Bioscience XF96 extracellular flux analyzer. After establishing a baseline, oligomycin (10 μM), FCCP (15 μM), and rotenone/antimycin A (1 μM, and 10 μM, respectively) were added. (G) Fold change in ATP levels in EV or shFLCN RAW264.7 after 24 h of seeding as measured by CellTiter-Glo Luminescent Cell Viability Assay. Data represent the average of three independent experiments, each done in triplicates ± SEM. Significance was determined using student’s t-test (**p<0.01, ***p<0.001). (H) Immunoblot analysis of EV and TFEB/TFE3 DKO RAW264.7 cells transfected with EV or shFLCN. (I) Phagocytic activities of EV, TFEB/TFE3 DKO, and TFEB/TFE3 DKO shFLCN RAW264.7 cells measured using Red pHrodo S.aureus BioParticles by flow cytometry. Data represents the average of three independent experiments, each done in triplicates ± SEM. Significance was determined using one-way ANOVA with the application of Bonferroni correction (**p<0.01, ****p<0.0001).