- Title
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Blockade of MCU-Mediated Ca2+ Uptake Perturbs Lipid Metabolism via PP4-Dependent AMPK Dephosphorylation
- Authors
- Tomar, D., Jaña, F., Dong, Z., Quinn, W.J., Jadiya, P., Breves, S.L., Daw, C.C., Srikantan, S., Shanmughapriya, S., Nemani, N., Carvalho, E., Tripathi, A., Worth, A.M., Zhang, X., Razmpour, R., Seelam, A., Rhode, S., Mehta, A.V., Murray, M., Slade, D., Ramirez, S.H., Mishra, P., Gerhard, G.S., Caplan, J., Norton, L., Sharma, K., Rajan, S., Balciunas, D., Wijesinghe, D.S., Ahima, R.S., Baur, J.A., Madesh, M.
- Source
- Full text @ Cell Rep.
Loss of MCU Limits mCa2+ Uptake and Elevates Total Body Fat in Zebrafish (A) Schematic of the generation of global MCU KO zebrafish using CRISPR/Cas9. (B) Genotyping for MCU deletion. (C) Bar graph showing MCU mRNA abundance in WT, MCU+/−, and MCU−/− zebrafish. (D) Western blot for MCU expression. n = 3. (E) Measurement of mCa2+ uptake. n = 5–10. (F) Quantification of ionomycin-induced peak mCa2+ levels from (E). n = 5–10. (G) Bar graph representing cellular ATP levels in cells isolated from MCU+/+, MCU+/−, and MCU−/− zebrafish. n = 4. (H) Adult WT and MCU−/− zebrafish were homogenized and centrifuged. MCU−/− zebrafish samples show a clear yellow color lipid layer on top of the protein lysate. n = 6. (I) Adult WT and MCU−/− zebrafish were stained with Lipid Green to monitor the distribution of lipids in the whole body. n = 3. (J and K) Bar graphs represent the quantification of Lipid Green staining from the dorsal (J) or tail (K) fin. n = 3. Statistical analysis: mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. |