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. 2017 Feb 4;18(2):322. doi: 10.3390/ijms18020322

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© 2017 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Suppressive effect of hydroethanolic extract of A. capillaris and three bioactive compounds on tumor necrosis factor receptor-associated factor 6 (TRAF6) and vacuolar H⁺-adenosine triphosphatase (V-ATPase) expression and their interaction. (A) The effect of A. capillaris hydroethanolic extract (ACHE) on TRAF6 and V-ATPase expression and (B) their interaction. (C) The effect of three bioactive compounds on TRAF6 and V-ATPase expression and (D) their interaction. RAW 264.7 cells were exposed to RANKL (50 ng/mL) for 5 days in the absence and presence of ACHE (20 μg/mL) or three bioactive compounds (10 μM). The cells were lysed and cell lysates were incubated with TRAF6 or V-ATPase antibody and protein A beads. After washing the beads, precipitated proteins and cell lysates were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotted for the indicated proteins using specific antibodies. Data are expressed as percentages of the values of untreated cells (means ± standard deviations, n = 3). ^## p < 0.01, ^### p < 0.001 vs. C; *** p < 0.001 vs. TC. C: control, which was not treated; TC: treated control, which was treated with RANKL; CA: chlorogenic acid; HP: hyperoside; SP: scoparone; NS: not significant.

Suppressive effect of hydroethanolic extract of A. capillaris and three bioactive compounds on tumor necrosis factor receptor-associated factor 6 (TRAF6) and vacuolar H⁺-adenosine triphosphatase (V-ATPase) expression and their interaction. (A) The effect of A. capillaris hydroethanolic extract (ACHE) on TRAF6 and V-ATPase expression and (B) their interaction. (C) The effect of three bioactive compounds on TRAF6 and V-ATPase expression and (D) their interaction. RAW 264.7 cells were exposed to RANKL (50 ng/mL) for 5 days in the absence and presence of ACHE (20 μg/mL) or three bioactive compounds (10 μM). The cells were lysed and cell lysates were incubated with TRAF6 or V-ATPase antibody and protein A beads. After washing the beads, precipitated proteins and cell lysates were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotted for the indicated proteins using specific antibodies. Data are expressed as percentages of the values of untreated cells (means ± standard deviations, n = 3). ^## p < 0.01, ^### p < 0.001 vs. C; *** p < 0.001 vs. TC. C: control, which was not treated; TC: treated control, which was treated with RANKL; CA: chlorogenic acid; HP: hyperoside; SP: scoparone; NS: not significant.