Figure 6
Effect of glutamine treatment on PfHz-mediated inhibition of HSP70. (A) Peripheral blood mononuclear cells (2.5 × 106) were incubated with LPS for 1 h prior to addition of Gln (0.5, 2.0 and 4.0 mM, pretreated 30 min prior to adding PfHz) for different time points. Total RNA was isolated and gene expression analysis by qRT-PCR was performed to quantify the HSP70 (1A + 1B) transcripts, using TaqMan® gene expression assays. β-actin was used as an endogenous control to normalize the data by the ΔΔCT method. Data represent average of individuals (n = 3) with each condition performed in triplicate (error bars represent SEM). * indicates P < 0.05 (one-way ANOVA) relative to LPS + PfHz treated cells. (B) Experiments performed as outlined in A showing the effect of Gln (0.5, 2.0 and 4.0 mM) on PfHz-mediated inhibition of Hsp70 assessed using immunoblotting at 24 h. (C) Densitometry analysis was performed to quantify intensity of expression. Data shown after normalizing Hsp70 relative to β-actin (n = 3); error bars represent SEM.