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. 2014 Jun 11;2(6):e12018. doi: 10.14814/phy2.12018

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© 2014 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — The effect of cell differentiation, OA, MO, lecithin, incubation time, and collagen matrix on lipoprotein secretion. (A) Cell differentiation: cells that were 4, 13, 17, or 23 days postconfluent were incubated for 4 h with 4 mmol/L OA, 1 mmol/L MO, 0.68 mmol/L lecithin, and 1 mmol/L NaTC in growth media (n = 4 for each; except for 13 days, n = 3). (B) OA: Thirteen‐day postconfluent cells were incubated for 4 h with 0, 2, 4, or 6 mmol/L OA in growth media containing 1 mmol/L MO, 0.68 mmol/L lecithin, and 1 mmol/L NaTC (n = 4 for 0 and 4 mmol/L; n = 5 for 2 mmol/L; n = 3 for 6 mmol/L). (C) MO: incubated with 0, 1, 2, or 4 mmol/L MO in growth media containing 2 mmol/L OA, 0.68 mmol/L lecithin, and 1 mmol/L NaTC (n = 5 for 0 and 4 mmol/L; n = 3 for 1 mmol/L; n = 7 for 2 mmol/L). (D) lecithin: incubated with 0, 0.68, 1.36, or 2.72 mmol/L lecithin in growth media containing 2 mmol/L OA and 1 mmol/L NaTC (n = 3 for 0 and 0.68 mmol/L; n = 4 for 1.36 and 2.72 mmol/L). (E) Time: incubated with 2 mmol/L OA, 1.36 mmol/L lecithin, and 1 mmol/L NaTC in growth media for either 2 or 4 h; and collected for either 2 or 4 h in growth media (n = 3 for 2 + 2 and 4 + 2; n = 4 for 2 + 4 and 4 + 4). (F) Collagen: Each 10 cm tissue culture dish was coated with 0, 50, 100 or 500 μg collagen before it was populated with cells (n = 3 for each; except 0 μg, n = 4). After the cells have reached 13‐day postconfluence, they were incubated for 4 h with 2 mmol/L OA, 1.36 mmol/L lecithin, and 1 mmol/L NaTC in growth media. The secreted lipoproteins were then collected for 2 h in growth media, and separated into CM and VLDL layers by sequential NaCl density gradient ultracentrifugation. The concentration of TG in the CM and VLDL layers was measured. Refer to Table 1 for the schematic representation of the experimental conditions. There were at least three separate experiments performed for each study. n represents the total number of dishes. All of the TG measurements were done in duplicates. Mean ± SE. One‐way ANOVA with multiple comparison tests was used (*P <0.05 when compared with the groups without *. Comparisons were made only among the CM samples; among the VLDL samples; not between the CM and the VLDL samples).