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. 2022 Dec 15;132(24):e157302. doi: 10.1172/JCI157302

Figure 4. Fatty acid is a negative feedback signal for viperin induction.

Figure 4

(A) MKN28 cells were cultured in serum-free media and then replenished with serum for the indicated durations. Viperin was detected by immunoblot using MaP.VIP. Grp94 was used as a loading control. (B) MKN28 was cultured in the conditioned media, RPMI complete media, serum-free RPMI or DMEM media, DMEM/F12 media, or RPMI supplemented with B27 for 48 hours. Viperin was detected by immunoblotting using MaP.VIP. Grp94 was used as a loading control. (C) Linoleic acid, a component of serum, is a key regulator of viperin induction in cancer cells. MKN28 cells were cultured in serum-free media and treated with linoleic acid–BSA or putrescine for 48 hours. Viperin was detected by immunoblotting using MaP.VIP. α-Tubulin was used as a loading control. (D) MKN28 cells were cultured in the presence and absence of serum or phenol red for 48 hours. Viperin was detected by immunoblotting using MaP.VIP. Grp94 was used as a loading control. (E) Fatty acid is a negative feedback signal for viperin induction. MKN28 cells were cultured in serum-free media and treated with linoleic acid–BSA, oleic acid–BSA, or palmitic acid–BSA for 48 hours. Viperin was detected by immunoblotting using MaP.VIP. Grp94 was used as a loading control. (F) MKN28 cells were cultured in serum-free media and treated with oleic acid for 48 hours. Viperin and HIF-1α were detected by immunoblotting using specific mAbs. Grp94 was used as a loading control.