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Figure 4 | Retrovirology

Figure 4

From: Glyceraldehyde 3-phosphate dehydrogenase negatively regulates human immunodeficiency virus type 1 infection

Figure 4

Effects of enhanced GAPDH packaging inside virions on HIV-1 replication. (A) Enhanced GAPDH packaging inside virions. The packaging efficiency of GAPDH was analyzed in lysates from viral particles produced from HEK293 cells cotransfected with pNL-CH and either the GAPDH expression or control vector. (B) Infectivity of enhanced-GAPDH-packaging virus. TZM-bl cells were incubated with an equal amount (2 ng of p24 antigen) of the enhanced-GAPDH-packaging virus or the control virus. Infectivity was evaluated as described in “Methods”. The mean values of at least three independent experiments are shown. (C) Quantification of tRNALys3 in enhanced-GAPDH-packaging virus. Incorporated tRNALys3 level was determined by reverse transcription quantitative PCR analysis as described in “Methods” and normalized by viral genomic RNA level. The amount of tRNALys3 in the control virus was set as 100%. The mean values of at least three independent experiments are shown. (D) Enhanced GAPDH packaging inside virions decreased reverse transcription efficiency in PMBCs. PBMCs were incubated with an equal amount (20 ng of p24 antigen) of the enhanced-GAPDH-packaging virus or the control virus. Reverse transcription products (R/gag DNA products) were determined as described in “Methods”. The significance of difference (Student’s t-test) is indicated as follows: **, p<0.05. The mean values of at least three independent experiments are shown. The error bars denote the standard deviation.

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