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LEGEND SUPPLEMENTARY FIGURES
Supplementary Figure 1. Plasma concentration of amino acids. A, concentration of essential
amino acids. B, concentration of non-essential amino acids. Wild-type and Mstn-/- mice were
inoculated with Lewis lung carcinoma (LLC) cells or received DPBS. Blood samples were
removed 35 days after the inoculation of LLC cells. Data are means ± SE (n=8-12/group). * P
< 0.05 and ** P < 0.01.
Supplementary Figure 2. Mstn gene inactivation prevents loss of skeletal muscle in tumorbearing mice. A, extensor digitorum longus muscle weight. B, soleus muscle weight. C,
tibialis anterior muscle weight. Wild-type and Mstn-/- mice were inoculated with Lewis lung
carcinoma (LLC) cells or received DPBS. Skeletal muscles were removed and weighted 35
days after the inoculation of LLC cells. Data are means ± SE (n=8-12/group). ** P < 0.01 and
*** P < 0.001.
Supplementary Figure 3. Wild-type and Mstn-/- tumor-bearing mice have similar caloric
intakes, indicating that anorexia is not involved in the development of cancer cachexia. Wildtype and Mstn-/- mice were inoculated with Lewis lung carcinoma (LLC) cells or received
DPBS. Food intake was normalized to tumor-free body weight. Data are means ± SE (n=812/group).
Supplementary Figure 4. Effect of human recombinant Mstn (hrMstn) on the proliferation
of Lewis lung carcinoma (LLC) cells - Bcl-2 protein content in the tumor tissue. A-C, number
(A), relative distribution of LLC cells in G1, S and G2M phases (B) and Ki-67 protein content
(C) of LLC cells cultured for 48 hours in absence and presence (2.5 μg.ml-1) of hrMstn.
Geometric means of the fluorescence intensity peaks were used to determine the expression of
Ki-67. Data are means ± SE (n=5-6 independent experiments). D, immunoblot analysis of
Bcl-2 protein content in the tumors of wild-type and Mstn-/- mice. Wild-type and Mstn-/- mice
were inoculated with Lewis lung carcinoma (LLC) cells. Tumor tissues were removed 35
days after the inoculation of LLC cells. Data are means ± SE (n=8-12/group).
Supplementary Figure 5. Lifespan of wild-type and Mstn-/- mice - Number of polyps. A,
Kaplan Meier plot illustrating the lifespan of wild-type (n=14) and Mstn-/- mice (n=14). B,
number of intestinal polyps according to their anatomical distribution (upper, middle and
lower intestine, colon) in ApcMin/+Mstn+/+ and ApcMin/+Mstn-/- mice. Mice were 23.3 ± 0.9
weeks old at the time of analysis. Data are means ± SE (n=4-7/group). * P < 0.05 and ** P <
0.01.
Supplementary Figure 6. Serum levels of interferon (IFN)-, interleukin (IL)-1, IL-6 and
tumor necrosis (TNF)-α in wild-type (n=7) and Mstn-/- mice (n=8). Data are means ± SE. * P
< 0.05.