Figure 2 | Scientific Reports

Figure 2

From: Nicotinamide N-methyltransferase (NNMT) regulates the glucocorticoid signaling pathway during the early phase of adipogenesis

Figure 2The alternative text for this image may have been generated using AI.

Nnmt deficiency impairs adipocyte differentiation by regulating MCE. (A) Terminal adipogenesis was analyzed 10 days after DIM induction in 3T3-L1-Nnmt-KO (3T3-L1-CRISPR/CAS9 Nnmt guides) and mock control cells (3T3-L1-CRISPR/CAS9 empty) by Oil Red-O staining and bright field microscopy (20× magnification). (B) Statistical data for Red-Oil staining quantification by OD measurements. Data are presented as means ± S.D. (n = 3) and statistical significance was calculated with a one-sided Welch’s t-test. (C) DNA content was analyzed by PI staining and FACS analysis was performed at different times after hormonal stimulation (T0, 20 h DIM and 48 h DIM). The percentage of cells in each phase of the cell cycle are represented in the graph plot, which shows the results of three independent experiments. Statistical significance was calculated by a chi-squared test using a 3 × 2 contingency table including the percentage of cells at each cell cycle stage (G1, S, G2/M) and in each experimental condition (mock – Nnmt-KO) per triplicate. (D) Flow cytometry plots representative of one independent experiment. (E) Annexin and PI staining. Nnmt-KO and mock cell lines were stained 48 h after DIM induction with Fluor 488 annexin V and PI and analyzed by flow cytometry. The loss of Nnmt did not influence either cell viability or apoptotic rate. For all the panels: *p < 0.05; **p < 0.01; ***p < 0.001.

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