FIG. 4.

Viable cell numbers, ATP contents, and media LDH levels after AFN treatment of mtCCs and exposure to RA or hyperoxia for 24 h in the presence or absence of BSO. mtCCs were treated with 1 μM AFN for 1 h, media changed, and subsequently exposed to RA or 85% O₂ for 24 h in the presence or absence of 0.25 mM BSO. (A) The addition of 0.25 mM BSO to the culture medium of AFN-treated cells decreased viable cell numbers in both RA and hyperoxia, as assessed by trypan blue exclusion. (B) ATP contents were lower in AFN-treated cells cultured in RA and hyperoxia in the presence of BSO. (C) Media LDH levels were greater in AFN-treated mtCCs cultured in RA or hyperoxia than in vehicle-treated mtCCs cultured under similar conditions. Data (mean±SEM, n=9–12) were analyzed by Kruskal–Wallis followed by Dunn's post hoc with significance noted at p<0.05 (*different than vehicle/RA; ^#different than vehicle/O₂; ^¥different than same treatment/RA; ^$different than AFN/O₂; ^¢different than AFN/RA; ^€different than BSO/RA). BSO, buthionine sulfoximine; LDH, lactate dehydrogenase.