Figure_7.tif (3.76 MB)

Blockade of the mevalonate pathway increases CD9 and CD81.

figure

posted on 2013-09-09, 02:54 authored by Yingji Jin, Isao Tachibana, Yoshito Takeda, Ping He, Sujin Kang, Mayumi Suzuki, Hanako Kuhara, Satoshi Tetsumoto, Kazuyuki Tsujino, Toshiyuki Minami, Takeo Iwasaki, Kaori Nakanishi, Satoshi Kohmo, Haruhiko Hirata, Ryo Takahashi, Koji Inoue, Izumi Nagatomo, Hiroshi Kida, Takashi Kijima, Mari Ito, Hideyuki Saya, Atsushi Kumanogoh

(A) RAW264.7 cells were untreated (-) or treated for 48 h with 50 ng/ml TSA (+) in the absence (-) or presence of 50 µM theophylline or 0.5 µM fluvastatin (Fluv) (+). The cells were lysed, and levels of CD9 and CD81 were examined by immunoblotting. Anti-actin blots show that comparable amounts of protein were loaded in each lane. (B) The mevalonate pathway and inhibitors. n-BP, nitrogenous bisphosphonate. (C) RAW264.7 cells were cultured for 24 h in the presence of indicated concentrations of fluvastatin, simvastatin (Simv), zoledronate (Zol), or risedronate (Ris). Levels of CD9 and CD81 were examined by immunoblotting. (D) RAW264.7 cells were cultured for 24 h in the absence (V, vehicle alone) or presence of mevalonate (Mev), farnesyl pyrophosphate (FPP), squalene (Squ), or geranylgeranyl pyrophosphate (GGPP). Although the actin level in the GGPP lane appears to be lower, an equal amount of protein was loaded. (E) RAW264.7 cells were cultured for 24 h in the absence (V) or presence of fluvastatin, zoledronate, farnesyl transferase inhibitor (FTI), or geranylgeranyl transferase inhibitor (GGTI). (F) RAW264.7 cells were untreated (-) or treated with fluvastatin (+) in the absence (V) or presence of mevalonate, FPP, squalene, or GGPP. (G) RAW264.7 cells were untreated (-) or treated with zoledronate (+) in the absence (V) or presence of mevalonate, FPP, squalene, or GGPP. (H) RAW264.7 cells were untreated (-) or treated with fluvastatin (+) in the absence (V) or presence of mevalonate, FPP, squalene, or GGPP and stimulated for 15 min with 0.1 µg/ml LPS (+). The cells were lysed, and levels of IκBα were examined by immunoblotting. (I) RAW264.7 cells were cultured for 24 h in the indicated concentrations of HA1077. Levels of CD9 and CD81 were examined by immunoblotting.