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. 2019 Feb 8;14(2):e0211954. doi: 10.1371/journal.pone.0211954

Fig 2. siRNA-mediated target gene knockdown capability was maintained using lipoplexes stored at different temperatures over time in vitro.

Fig 2

(A) HeLa cells were treated with a final concentration of 40 nM of siGlo complexed into lipoplexes and incubated at 37°C for 24h before assessing siGlo uptake (%) by flow cytometry. Tests were performed on freshly prepared lipoplexes (F–Black shaded bars), and lipoplexes stored at various temperatures for different lengths of time post manufacture. Data is presented as percentage sum of fluorescence intensity. Bar graphs represent the mean and the error bars represent the standard deviation from three independent experiments. (B) HeLa cells were treated with a final concentration of 40 nM of Lamin A/C (LMNA) siRNA complexed into lipoplexes and incubated at 37°C for 3 days. Expression levels were calculated relative to non-transfected cells (control) by qPCR analysis and data presented as percentage of LMNA expression compared to control. β-actin was used as housekeeping control gene. (C) Percentage of knockdown efficiency was also determined as previously described [21]. Tests were performed on freshly prepared lipoplexes (F–Black shaded bars), and lipoplexes stored at various temperatures for different lengths of time post manufacture. Bar graphs represent the mean and the error bars represent the standard deviation from three independent experiments. **p value < 0.005, ***p value < 0.0005, ****p value <0.0001, two-way ANOVA using a Dunnett's post-hoc analysis against freshly prepared lipoplexes.