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. 2024 Sep 18;16(18):3146. doi: 10.3390/nu16183146

Figure 1.

Figure 1

Evaluation of gene expression in hepatocytes after transfection with plant miR6262 mimic and treatment with free fatty acids to mimic in vitro liver steatosis. (a) mRNA expression of the predicted target RXRA. (b) mRNA expression of transcription factors (PPARA, FOXO1, and SREBF1) and functional protein-coding genes (MAPKAPK2, QKI, FASN, ACOX1, G6PC, and GSK3B) associated with lipid and glucose metabolism. Reverse-transfection of HepG2 cells was conducted with the plant miR6262 mirVana mimic (5′-UCUUUAGAAAGUUAGAAUUGU-3′) and a negative control (a scramble sequence) at 50 nM. At 48 h post-transfection, cells were exposed to free fatty acids (a mixture of oleic and palmitic acids at 0.5 M and a 2:1 ratio) for 3 h. Quantification of mRNA expression levels was evaluated by qPCR. TBP was selected as housekeeping gene to normalize Cq values, which were expressed relative to the free FFA-treated negative control, applying the 2−ΔΔCt method. Results are shown as relative gene expression mean relative gene expression mean ± standard error of the mean (SEM) (n = 4–5). p-value: * p < 0.05, ** p < 0.01, *** p < 0.001. Abbreviations: NC (negative control), FFA (free fatty acids).