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Tomar et al., 2019 - Blockade of MCU-Mediated Ca2+ Uptake Perturbs Lipid Metabolism via PP4-Dependent AMPK Dephosphorylation. Cell Reports   26:3709-3725.e7 Full text @ Cell Rep.

Fig. 3

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.

Acknowledgments:
ZFIN wishes to thank the journal Cell Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Rep.