Fig. 3. Coculture supernatant induces iNOS in astrocytes. Total RNA was extracted from MDM/HFA cocultures at day 18 postinfection and from HFA cultures incubated for 7 days with different supernatants. Levels of iNOS mRNA in HFA were analyzed by RT-PCR. The cytokine-stimulated A172 cell line was used as a positive control. Lane 1, positive control; lane 2, HFA; lane 3, HFA cocultured with uninfected MDM; lane 4, HFA cocultured with infected MDM; lane 5, HFA incubated with HIV-1; lane 6, HFA incubated with day 18 to 21 supernatant of uninfected MDM culture; lane 7, HFA incubated with day 18 to 21 supernatant of infected MDM culture; lane 8, HFA incubated with day 18 to 21 supernatant of coculture with uninfected MDM and HFA; lane 9, HFA incubated with day 3 to 6 supernatant of coculture with infected MDM and HFA; lane 10, HFA incubated with day 18 to 21 supernatant of coculture with infected MDM and HFA.
Fig. 3.

Coculture supernatant induces iNOS in astrocytes. Total RNA was extracted from MDM/HFA cocultures at day 18 postinfection and from HFA cultures incubated for 7 days with different supernatants. Levels of iNOS mRNA in HFA were analyzed by RT-PCR. The cytokine-stimulated A172 cell line was used as a positive control. Lane 1, positive control; lane 2, HFA; lane 3, HFA cocultured with uninfected MDM; lane 4, HFA cocultured with infected MDM; lane 5, HFA incubated with HIV-1; lane 6, HFA incubated with day 18 to 21 supernatant of uninfected MDM culture; lane 7, HFA incubated with day 18 to 21 supernatant of infected MDM culture; lane 8, HFA incubated with day 18 to 21 supernatant of coculture with uninfected MDM and HFA; lane 9, HFA incubated with day 3 to 6 supernatant of coculture with infected MDM and HFA; lane 10, HFA incubated with day 18 to 21 supernatant of coculture with infected MDM and HFA.

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