Author: Ranjbar, Shahin; Haridas, Viraga; Jasenosky, Luke D.; Falvo, James V.; Goldfeld, Anne E.
Title: A Role for IFITM Proteins in Restriction of Mycobacterium tuberculosis Infection Document date: 2015_10_22
ID: zy82bo7q_40
Snippet: THP-1 cells expressing the IFITMs' shRNA or control shRNA (1 × 10 6 ) were seeded in sixwell plates in triplicate and infected with H37Rv. After a 3-hr incubation at 37°C, the cultures were treated with 50 μg/ml puromycin for 20 min. Cells were washed three times with PBS; cultured in 3 ml fresh RPMI plus 10% FCS for 0, 48, and 72 hr; and incubated at 37°C and 5% CO 2 . Following each time point, cultures were terminated, cells were treated w.....
Document: THP-1 cells expressing the IFITMs' shRNA or control shRNA (1 × 10 6 ) were seeded in sixwell plates in triplicate and infected with H37Rv. After a 3-hr incubation at 37°C, the cultures were treated with 50 μg/ml puromycin for 20 min. Cells were washed three times with PBS; cultured in 3 ml fresh RPMI plus 10% FCS for 0, 48, and 72 hr; and incubated at 37°C and 5% CO 2 . Following each time point, cultures were terminated, cells were treated with lysis buffer and sonicated, and serial dilutions were prepared and inoculated in Middlebrook 7H11 agar in triplicate. Colonies were analyzed on day 25 post-infection.
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