1. Volume 21, Issue 11, Pages 3129-3140 (December 2017)
Adipose KLF15 Controls Lipid Handling to Adapt to Nutrient Availability 
Keiichiro Matoba, Yuan Lu, Rongli Zhang, Eric R. Chen, Panjamaporn Sangwung, Benlian Wang, Domenick A. Prosdocimo, Mukesh K. Jain 
Cell Reports 
Volume 21, Issue 11, Pages (December 2017)
DOI: /j.celrep
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2. Cell Reports 2017 21, 3129-3140DOI: (10.1016/j.celrep.2017.11.032)
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3. Figure 1 Targeted Deletion of Adipose Klf15 Decreases Adiposity
(A) Recombination efficiency was determined in adipocytes isolated from epididymal WAT (eWAT), inguinal WAT (iWAT), liver, skeletal muscle, heart, and kidney by qPCR. n = 3.
(B) Body weight of control and AK15KO mice fed standard chow. n = 26–32.
(C) Fat and lean mass measured by magnetic resonance imaging. n = 3.
(D) Daily food intake monitored in individualized cage. n = 10.
(E) Gross morphology of whole body (upper panel) and fat pads (lower panel).
(F) Tissue weights. n = 6–7. BAT, brown adipose tissue.
(G) Representative images of H&E-stained sections of eWAT (upper panels) and iWAT (lower panels). Scale bars, 100 μm.
(H) Frequency distribution of adipocyte size of eWAT (upper panel) and iWAT (lower panel). n = 4.
(I) Mean adipocyte diameter of eWAT (upper panel) and iWAT (lower panel). n = 4.
(J) Acute adenoviral silencing (left panel) and overexpression (right panel) of Klf15 in differentiated 3T3-L1 cells. n = 3. EV, empty vector; sh-KLF15, shRNA targeting Klf15; Ad-KLF15, adenoviral Klf15 overexpression.
(K) Triglyceride (TG) content in the same cells. n = 3.
(L and M) Intravenous glucose tolerance tests (L) and insulin tolerance tests (M). n = 4. Bar charts show the area under the curve (AUC) of glucose.
(N) Akt activation in basal or insulin-stimulated skeletal muscle and liver. Bar chart showing the quantification of insulin-induced phosphorylated Akt to total Akt. p < Data are represented as mean ± SEM.
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4. Figure 2
KLF15 Regulates Genes Important for Lipid Synthesis in Adipocytes
(A) 2-DG6P uptake in 3T3-L1 cells under basal or insulin (1 μM)-stimulated conditions.
(B) Glycolysis determined by Flux Analyzer with 3T3-L1 adipocytes. Oligo, oligomycin (ATP synthase inhibitor). Bar chart showing the AUC of ECAR. n = 3.
(C) GLUT4 immunostaining in eWAT. Corresponding quantification is shown in the right panel. n = 4. Scale bars, 100 μm.
(D and E) The mRNA levels of lipogenesis-related genes determined by qPCR in adipocytes isolated from eWAT (D), or in 3T3-L1 cells after Klf15 knockdown (E, left) and overexpression (E, right). n = 3. p < Data are represented as mean ± SEM.
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5. Figure 3 Increased Lipolysis in Adipose-Specific Klf15 Knockout Mice
(A) Basal or isoproterenol (ISO, 1 μM)-stimulated release of glycerol and fatty acids from eWAT explants. n = 3.
(B) Medium levels of glycerol and fatty acids in basal or ISO (1 μM)-stimulated 3T3-L1 cells. n = 3.
(C) Serum levels of glycerol (left) and fatty acids (right) measured before and after insulin injection during the ITT (see Figure 1M). n = 3.
(D and E) Western blot of phosphorylated HSL and total HSL in eWAT (D) or 3T3-L1 adipocytes (E). Corresponding quantifications are shown in the right panels. p < 0.05 versus EV.
(F) Cellular cAMP levels in same cells. n = 3. p < 0.05 versus EV.
(G and H) Western blot of insulin signaling molecules in insulin (0.75 U/kg, 15 min)-injected mice eWAT (G) or insulin-stimulated (100 nM, 5 min) 3T3-L1 cells (H). Corresponding quantifications (insulin-stimulated conditions) are shown in the right panels. Ins, insulin.
(I and J) The mRNA levels of lipolysis-related genes were determined by qPCR in adipocytes isolated from eWAT (I), or in 3T3-L1 cells after Klf15 knockdown (J, left) and overexpression (J, right). n = 3. p < Data are represented as mean ± SEM.
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6. Figure 4
Adipose Klf15 Deficiency Enhances Muscle Performance and Liver Ketogenesis
(A) Change in serum levels of fatty acids and β-hydroxybutylate (β-HB) in wild-type mice during fed (black bar), 24 hr fasting (white bar), 24 hr fasting followed by 24 hr of refeeding (gray bar). n = 5. ∗p < 0.05 versus fed, #p < 0.05 versus fast state.
(B) Relative expression levels of Klf15 in adipocytes isolated from eWAT obtained from wild-type mice under same conditions. n = 5. ∗p < 0.05 versus fed, #p < 0.05 versus fast state.
(C) Time course analysis of Klf15 expression in white adipocytes obtained from fasted wild-type mice. n = 3. p < 0.05 versus time 0.
(D) Running time to exhaustion on an endurance exercise protocol under fed or overnight (16 hr) fasted conditions. n = 6.
(E) Serum ketone levels assessed by β-HB concentration under fed or overnight (16 hr) fasted conditions. n = 5–9. p < Data are represented as mean ± SEM.
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7. Figure 5
Adipose-Specific Klf15 Deletion Attenuates Fat Accumulation under High-Fat Diet Feeding
(A) Body weight of control and AK15KO mice fed HFD (41 kcal% fat). n = 11–18.
(B) Gross morphology of whole body (left) and fat pads (right).
(C) Fat tissue weights. n = 7.
(D–H) Representative images of H&E-stained sections (D), frequency distribution of adipocyte size (E), and mean adipocyte diameter (F) of eWAT obtained from HFD-fed mice. n = 4. Scale bars, 100 μm. Intraperitoneal glucose tolerance tests (G) and insulin tolerance tests (H). Bar charts show the AUC of glucose. n = 4–7.
(I and J) The mRNA levels of adipogenesis (I) or lipogenesis (J)-related genes determined by qPCR in adipocytes isolated from eWAT of mice with HFD. n = 6.
(K) Basal or isoproterenol (ISO, 1 μM)-stimulated release of glycerol and fatty acids from eWAT explants of HFD mice. n = 3.
(L) Western blot of insulin signaling molecules in HFD mice eWAT. Corresponding quantifications are shown in the right panels.
(M) The mRNA levels of lipolysis-related genes determined by qPCR in adipocytes isolated from eWAT. p < Data are represented as mean ± SEM.
Cell Reports  , DOI: ( /j.celrep )
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