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Additional Supporting Information may be found in the online version of this article.

FilenameFormatSizeDescription
sc-12-0136_sm_SupplFigure1.tif956KFigure S1. Overexpression of C/EBPβ in myoblasts does not affect cell growth. (A) Crystal violet assay to determine cell number of satellite cell cultures retrovirally transduced to express C/EBPβ or with empty vector (pLX) and cultured for 48 hours in growth medium. Error bars are the SEM, n=3. (B) Crystal violet assay to determine cell number of C2C12 cells retrovirally transduced to express C/EBPβ or with empty vector (pLX) and cultured for 48 hours in growth medium. Error bars are the SEM, n=4.
sc-12-0136_sm_SupplFigure2.pdf96KFigure S2. Restoration of MyoD protein expression rescues the differentiation defect in C/EBPβ-overexpressing C2C12 cells. (A) Indirect immunostaining for MHC expression in C2C12 cells retrovirally transduced to express C/EBPβ or with empty vector (pLXSN), and transiently transfected to express GFP and MyoD as indicated. Cells were differentiated in low serum conditions for 4 days prior to immunostaining. DAPI staining reveals nuclei. Scale bar = 50μm. (B) Differentiation index of cultures treated as in (A). *p<0.05, n=3. Error bars are the standard deviation. (C) Western analysis of C/EBPβ, MyoD and myogenin expression in C2C12 cultures transfected and differentiated as in (A) β-tubulin is used as a loading control. (D) Images of GFP expression, merged with phase contrast images, to determine transfection efficiency. (E) Transfection efficiencies, defined as the percentage of GFP+ cells in the culture 24 hours after transfection, for each transfection condition. One way ANOVA analysis revealed a significant decrease (p<0.05) between conditions labeled a and b.
sc-12-0136_sm_SupplFigure3.tif1080KFigure S3. Loss of MyoD expression is partially rescued by the inhibition of the proteasome. (A) Western analysis of C2C12 myoblasts retrovirally transduced to express C/EBPβ or with empty vector (pLXSN) and cultured in 2% horse serum for 36 hours. Two hours prior to harvest, cells were treated with 50 μM MG132. Cyclophilin B is used as a loading control. (B) Western analysis of C2C12 myoblasts retrovirally transduced to express C/EBPβ or with empty vector (pLXSN) and cultured in growth medium pretreated with MG132 for 2 hours prior to harvest.
sc-12-0136_sm_SupplFigure4.tif535KFigure S4. Withdrawal of HGF from growth medium reduces Cebpb expression and promotes differentiation. (A) RT-qPCR analysis of Cebpb, Pax7 and Myog expression in WT satellite cell cultures grown in complete growth medium or growth medium lacking HGF for 48 hours. *p<0.05, **p<0.01, n=4. (B) Western analysis of C/EBPβ and Pax7 expression in SCs cultured and treated as in (A).
sc-12-0136_sm_SupplFigure5.pdf113KFigure S5. Fiber hypertrophy increased at day P56 in conditional knockout mice. (A) Representative bright field images of tibialis anterior cross sections from control Cebpbfl/fl and conditional null Cebpb-/-Pax7Cre/+ animals at postnatal day 56 stained with hematoxylin and eosin. Scale bar = 100 μm. (B) Average cross-sectional areas of muscle fibers from control and conditional null animals. For each group n=2 animals (1 male and 1 female). Error bars are the standard deviation. (C) Relative fiber number in the tibialis anterior muscles of control and conditional null mice (n=2 for each group). Control animals are set arbitrarily at 100. (D) Frequency distribution of fiber size in control and conditional null mice. Error bars are the standard deviation.
sc-12-0136_sm_SupplMethods.pdf76KSupplementary Data

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