Figure 2.
Quantification of intracellular lipid levels in hepatocytes after transfection with plant miR6262 mimic. (a) Intracellular triglycerides levels measured by Nile Red staining. (b) Intracellular triglycerides levels measured by Triglyceride-GloTM Assay. 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. Scramble-sequence-transfected cells, untreated with FFA, served as positive control. Results are expressed as the percentage of lipid accumulation, reflecting the mean ± standard error of the mean (SEM) of the lipid content versus to FFA-treated negative control (Nile red staining: n = 6–7; Triglyceride-GloTM assay: n = 3). Statistical significance was assessed by comparing miR6262 transfected cells and untreated negative control to the FFA-treated negative control. p-value ** p < 0.01, *** p < 0.001. Abbreviations: NC (negative control), FFA (free fatty acids).