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. 2019 Feb 1;30(3):387–399. doi: 10.1091/mbc.E18-09-0548

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© 2019 Melville et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 6: — FABP5 affects collagen budding and secretion. (A) Immunoblot of response of traditional COPII cargoes ERGIC53 and SEC22B to FABP5 added to budding reactions with HeLa donor membranes. Ribophorin used as a marker of ER contamination. (B) Schematic of OptiPrep gradient flotation of budding reaction using IMR90 cell donor membrane. (C) Immunoblot of cargoes from budding reaction, with ribophorin as a control for ER contamination. (D) Immunoblot of type I collagen in media and procollagen in cells after 30 min of ascorbic acid treatment in IMR90 and U2OS cells after siRNA knockdown of FABP5. Actin was used as loading control. (E) Quantification by Student’s two-tailed t test; p values indicated (n = 3). (F) Immunoblot of type I collagen in media and procollagen in cells after 30 min of ascorbic acid treatment in IMR90 after overnight doxycycline treatment to overexpress FABP5. Actin was used as loading control. (G) Quantification by Student’s two-tailed t test; p values indicated (n = 4). (H) Immunofluorescently labeled IMR90 cells. Nuclei are stained with DAPI. (I) Quantification of collagen and FABP5 levels observed in 100 cells.