Figure 2.

MTLN Regulates Blood Lipid Levels and Adipocyte Lipid Metabolism

(A) qRT-PCR analysis of 1500011K16Rik in primary mouse liver, brown adipose tissue (BAT), and gonadal adipose tissue (gWAT). n = 3 litter-matched mice, 3 technical replicates each.

(B–D) Serum triglycerides (B), total cholesterol (C), and free fatty acids (D) in mice fed a high-fat diet for 6 weeks, one matched cohort.

(E) Immunofluorescence imaging of murine pre-adipocytes stained for PPARγ2 (red) and nuclei (blue). Scale bars, 100 μm.

(F) High-content imaging quantification of the percentage of PPARγ2-positive nuclei in differentiated murine pre-adipocytes. n = 6.

(G) Quantification of triglyceride accumulation in differentiated murine pre-adipocytes. n = 16 for WT, 8 for KO, 7 for OE biological replicates from one differentiation.

(H) Concentration of glycerol released into the medium of differentiated murine pre-adipocytes under basal or forskolin-stimulated conditions. n = 6 for WT, 3 for KO, 3 for OE.

(I) Immunofluorescence imaging of genome-edited hPSC-adipocytes stained for C/EBP-α (CCAAT/enhancer binding protein alpha) (green), neutral lipids (red), and nuclei (blue). Scale bars, 100 μm.

(J) High-content imaging quantification of the percentage of C/EBP-α-positive nuclei in differentiated hPSC-adipocytes. n = 20 for WT, 40 for KO and OE.

(K) Quantification of triglyceride accumulation in hPSC-adipocytes. n = 33 for WT, 54 for KO, 75 for OE.

(L) Concentration of glycerol released into the medium of hPSC-adipocytes under basal or forskolin-stimulated conditions. n = 30/21 for WT, 42/42 for KO, 66/90 for OE basal/stimulated.

(M) Immunofluorescence imaging of genome-edited SGBS adipocytes stained for C/EBP-α (green), neutral lipids (red), and nuclei (blue). Scale bars, 500 μm.

(N) High-content imaging quantification of the percentage of C/EBP-α-positive nuclei in differentiated SGBS adipocytes. n = 9 for parental, GFP, and MTLN OE, 12 for MTLN KO and scrambled guide RNA control.

(O) Quantification of triglyceride accumulation in SGBS adipocytes. n = 9 for WT, 3 for KO, 12 for OE.

(P) Concentration of glycerol released into the medium of SGBS adipocytes under basal or forskolin-stimulated conditions. n = 3 for WT, KO, and OE.

Data are means ± SD. ^∗p ≤ 0.05, ^∗∗p ≤ 0.01, ^∗∗∗p ≤ 0.001, ^∗∗∗∗p ≤ 0.0001; one- or two-way ANOVA. Unless specified, “WT” groups include both untransduced and control vector overexpression cells. All n values are independent biological replicates, with 25 fields per replicate averaged for the imaging, and all lipid accumulation and glycerol release done in technical duplicate. Related to Figure S2.