Figure EV5. Lower expression levels of GDF11 induce loss of body weight and muscle atrophy.

1. Eleven‐week‐old C57BL/6 male mice received i.p. injections of PBS (control; n = 4), 1 × 10¹¹ gc of AAV8.GDF11 (mid dose; n = 3), or 5 × 10¹⁰ gc of AAV8.GDF11 (low dose; n = 3), and their body weights were monitored every other day until the 1 × 10¹¹ gc AAV8.GDF11 group required euthanasia on day 10.
2. Immunoblotting of serum samples for GDF11 demonstrates expression levels of 0.5–1.6 ng/μl and 0.26–0.43 ng/μl for the mid‐ and low‐dose groups, respectively.
3. Change in mouse body weights (Bwt) across the 10‐day study by the AAV8.GDF11 treatment groups, including terminal values for the previous 7‐day cohort treated with 1 × 10¹² gc of AAV8.GDF11 (high dose).
4. Mass of the quadriceps, gastrocnemius, and heart of the 10‐day study mice (n = 12 for control mice by the inclusion of untreated age‐matched mice, resulting in a larger, homogenous data set).
5. Liver and kidney mass of 7‐week‐old C57BL/6 male mice were treated with PBS (control; n = 5), AAV8.GDF11 low dose (n = 4), or AAV8.Mstn high dose (n = 5) for 16 days (see Fig 6B). The mean values for 7‐week‐old mice from this colony (n = 5) are indicated by the dotted line to show starting masses.
6. Immunoblotting comparison of monomeric GDF11 and Mstn levels in quadriceps, heart, and kidney demonstrate that differential effects are not due to differential accumulation of the ligands in tissue.

Data information: Values are displayed as mean ± SEM. Statistical analysis performed using one‐way ANOVA analysis with Tukey's HSD post hoc test (non‐connecting letters indicate P < 0.05 between groups) and effect size presented as eta‐squared (η²). Note that “b” on Day 4 of panel (C) refers to the 2 overlapping groups.