Journal: Infectious Agents and Cancer

Article Title: HIV Nef enhances the expression of oncogenic c-MYC and activation-induced cytidine deaminase in Burkitt lymphoma cells, promoting genomic instability

doi: 10.1186/s13027-020-00320-9

Figure Lengend Snippet: Fig. 1 AID and c-MYC expressions are altered in B cells upon exposure to HIV Nef. a. Bar graphs showing relative mRNA expression of c-MYC (left panel) and AICDA (right panel) at 3 h, 6 h and 12 h exposure to HIV Nef, in lymphoblastoid cells (LCLs) and Burkitt lymphoma cells (Ramos). Cells were treated at the indicated concentrations of recombinant Nef, total RNA was isolated and qPCR was performed on reverse transcribed mRNA using primers specific to c-MYC or AICDA. The levels were normalised to the internal control GAPDH and plotted relative to mock (0 ng/ml) treated cells. *** p ≤0.001; **p ≤0.01; * p ≤0.05. All experiments were performed in triplicate. Bars indicate standard deviation. b. Ramos cells were treated at the indicated concentrations of recombinant Nef, total protein was isolated, separated on 12% SDS-PAGE and analysed by western blotting with antibodies against c-MYC and AID. p38 was detected as a loading control. All experiments were performed in triplicate

Article Snippet: Real-time PCR was performed on a LightCycler® 480 (Roche, Germany) using KAPA SYBR® FAST qPCR Kit (Kapa Biosystems, South Africa).

Techniques: Expressing, Recombinant, Isolation, Reverse Transcription, Control, Standard Deviation, SDS Page, Western Blot