Figure 2

CDK2 KD inhibits HIV-1 transcription. (A) Expression of CDK2 mRNA is reduced in CDK2 KD cells. Total RNA was extracted from 293T cells and 293T-CDK KD cells lysates, reverse transcribed and analyzed by real-time PCR. β-Actin was used as an internal control. Quantification is shown in triplicates. (B) Cell cycle analysis of 293T-59 cells. Cells were fixed with 70% ethanol, stained with Propidium Iodine and analyze by FACS. Quantification is shown in triplicates. (C) CDK2 protein expression is reduced in 293T-CDK KD cells. Lysates from 293T and 293T-CDK2 KD cells (lanes 1 and 2) were analyzed for CDK2 expression by western blotting. Quantification is shown for three independent experiments. (D) Inhibition of VSVG HIV-1 Luc replication. 293T and 293T-CDK2-KD cells were infected with VSVG-pNL4-3 Luc virus and luciferase activity was measured. MTT assay was used for the normalization. Quantification is shown for three independent experiments. (E) CDK2 protein expression is reduced in HeLa-CDK2 KD cells. CDK2, tubulin and CDK9 expression was analyzed in HeLa and HeLa-CDK2 KD cells (lanes 1 and 2) by western blotting. (D) Inhibition of VSVG HIV-1 Luc replication. Lane1, CDK2 expression was analyzed in HeLa and HeLa-CDK2 KD cells by Real-time PCR. 18S RNA was used as an internal control. Quantification is shown in triplicates. Lane 2, Luciferase expression of VSVG-HIV-1 Luc was analyzed in infected HeLa and HeLa -CDK2-KD cells at 48 hours post infection. MTT assay was performed for the normalization. Quantification is shown for three independent experiments.