1. Volume 22, Issue 5, Pages 604-610 (May 2015)
Targeting Cholesterol in a Liquid-Disordered Environment by Theonellamides Modulates Cell Membrane Order and Cell Shape 
Yuko Arita, Shinichi Nishimura, Reiko Ishitsuka, Takuma Kishimoto, Junichi Ikenouchi, Kumiko Ishii, Masato Umeda, Shigeki Matsunaga, Toshihide Kobayashi, Minoru Yoshida 
Chemistry & Biology 
Volume 22, Issue 5, Pages (May 2015)
DOI: /j.chembiol
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2. Chemistry & Biology 2015 22, 604-610DOI: (10. 1016/j. chembiol. 2015
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3. Figure 1 Chemical Structures of TNM-A and TNM-AMCA
Chemistry & Biology  , DOI: ( /j.chembiol )
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4. Figure 2 Binding of TNM-AMCA to Cholesterol in the Ld Phase
(A) Cholesterol-dependent binding of TNM-AMCA to POPC-based GUVs.
(B) Binding of TNM-AMCA or EGFP-D4 to DOPC-based GUVs (filled) and SM-based GUVs (empty). GUV compositions were DOPC/Chol/Rh-DOPE (49:50:1) and SM/Chol/Rh-DOPE (49:50:1).
(C) Localization of TNM-AMCA in Ld domains. Confocal sections in the equatorial plane of a GUV (DOPC/SM/Chol/Rh-DOPE = 33:33:33:1) are shown. GUVs were incubated with 1 μM TNM-AMCA (blue) and EGFP-lysenin (green). Scale bar represents 10 μm.
(D) Effect of acyl chains on TNM-AMCA affinity. GUV composition was PC/Chol/Rh-DOPE (49:50:1). (A, B, and D) The fluorescence intensity of TNM-AMCA was normalized by that of Rh-DOPE. Data represent mean ± SD, n = 3 different GUVs.
Chemistry & Biology  , DOI: ( /j.chembiol )
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5. Figure 3 Phase Separation of Lipid Membranes by TNM-A
GUVs (DOPC/SM/Chol/Rh-DOPE, 24:25:50:1) were incubated with 1 μM TNM-A for 20 min.
(A) Confocal sections in the equatorial plane of GUVs are shown. Scale bar represents 10 μm.
(B) Number of GUVs in which phase separation was observed. Data represent mean ± SD, n = 6 independent experiments (n > 100 for each experiment).
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6. Figure 4 Effect of TNM-A on the Membrane Order
Cells pre-incubated with di-4-ANEPPDHQ (2 μM) were treated with TNM-A (0.5 μM), and pseudocolored GP value images of live cells were obtained. The red regions indicate higher order (Lo) membrane domains, while blue regions indicate lower order (Ld) domains. Scale bar represents 50 μm.
Chemistry & Biology  , DOI: ( /j.chembiol )
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7. Figure 5 Cell Contraction after Being Challenged with TNM-A
(A) Transient contraction of A549 cells by TNM-A. Cells were treated with 0.5 μM TNM-A in serum-free medium for 0.5 or 12 hr.
(B) Cells pre-treated with cytoskeleton modulators were exposed to TNM-A (1 μM) for 0.5 hr. Cells were fixed, stained with BODIPY-FL-phallacidin (green), anti-α-tubulin mAb (red), and Hoechst (blue, only in B). Scale bars represent 15 μm.
Chemistry & Biology  , DOI: ( /j.chembiol )
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