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. 2024 Sep 23;16(9):1238. doi: 10.3390/pharmaceutics16091238

Figure 4.

Figure 4

Analysis of transfection-mediated intracellular CDK8-specific siRNA knock-down (KD) in primary fibroblasts. (A) The CDK8-specific siRNA KD was specifically reduced, under slightly optimized conditions (Lipofectamine™ RNAiMAX), using the siRNAs CDK8 100 at a final concentration of 10 pmol/well. HFFs were cultivated in 6-well plates and infected with HCMV AD169 at MOI of 0.1 at two d post-siRNA transfection (see KD effect highlighted by dashed box). As control samples, mock-infected cells, mock-transfected (no siRNA) cells, a non-specific siRNA (scrambled), and a GAPDH-specific siRNA were used. Cells were harvested at 7 d p.i. for the preparation of total lysates, to be analyzed by SDS-PAGE/Wb procedures. (B) Densitometric analysis was performed in quadruplicate measurements (SDS-PAGE/Wbs in duplicate, densitometry in duplicate) using AIDA image analyzer, and statistical evaluation was conducted using ANOVA followed by Bonferroni’s correction method for multiple comparison (** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001).