Fig 1. Establishment of a neurons-hμglia/HIV co-culture system.

The time line is indicated by the large black arrow, with the corresponding day number shown in red. Undifferentiated, neuron precursor cells LUHMES/RFP are plated and allowed to expand for 2 days prior to transferring to the experimental wells at 500,000 neurons per well of a six-well plate at day 0. After 2 days, the neuronal differentiation is initiated by adding modified neuronal differentiation medium (mNDM). After differentiation for 1 day, 60,000 immortalized human microglial cells (hμglia/HIV) carrying a single round HIV construct with a GFP reporter (diagrammed, bottom left) are added. The viability of the neurons and the expression levels of HIV are monitored for 3 days.