Fig 1: ActE signals via the activin type II receptors.(A) Schematic diagram of activin type II receptor Fc-fusion proteins used as decoy receptors. (B and C) (CAGA12)-luciferase HEK293T293 cells transfected with ALK4-ST or ALK7-ST receptor constructs and treated with ActA (0.62nM), ActE conditioned media (20x), ActC (0.62nM), and 10μM SB-431542 in the presence of increasing amounts (6.25, 12.5, 25 nM) of either ActRIIA-Fc (****P < 0.0001) (****P < 0.0001) (****P = 0.0002) (B) or ActRIIB-Fc (****P < 0.0001) (P = 0.3132) (P = 0.0549) (ns = not significant) (C). (D) Schematic representation of neutralizing antibodies targeting the extracellular domains (ECDs) of the activin type II receptors. (E) (CAGA12)-luciferase HEK293T293 cells either untransfected or transfected with the ALK7-ST type I receptor and treated with ActA (0.62nM), ActE conditioned media (20x), ActC (0.62nM), and 10μM SB-431542 in the presence or absence of neutralizing antibodies targeting ActRIIA, ActRIIB, or both (2.5μg/mL) (****P < 0.0001) (****P < 0.0001) (****P < 0.0001) (ns = not significant) . In (B, C, and E) each point represents a technical replicate within triplicate experiments with bars displaying the mean ± SD. Data are represented as percentages of uninhibited signal (B, C, and E).
Fig 2: Activin E signals via the activin type II receptors.(A) Schematic diagram of activin type II receptor Fc-fusion proteins used as decoy receptors. (B and C) (CAGA12)-luciferase HEK293T293 cells transfected with ALK4-ST or ALK7-ST receptor constructs and treated with ActA (0.62 nM), ActE conditioned media (20×), ActC (0.62 nM), and 10 μM SB-431542 in the presence of increasing amounts (6.25, 12.5, 25 nM) of either ActRIIA-Fc (****P < 0.0001) (****P < 0.0001) (****P = 0.0002) (B) or ActRIIB-Fc (****P < 0.0001) (P = 0.3132) (P = 0.0549) (ns = not significant) (C). (D) Schematic representation of neutralizing antibodies targeting the extracellular domains (ECDs) of the activin type II receptors. (E) (CAGA12)-luciferase HEK293T293 cells either untransfected or transfected with the ALK7-ST type I receptor and treated with ActA (0.62 nM), ActE conditioned media (20×), ActC (0.62 nM), and 10 μM SB-431542 in the presence or absence of neutralizing antibodies targeting ActRIIA, ActRIIB, or both (2.5 µg/ml) (****P < 0.0001) (****P < 0.0001) (****P < 0.0001) (ns = not significant). In (B, C, and E) each point represents a technical replicate within triplicate experiments with bars displaying the mean ± SD. Data are represented as percentages of uninhibited signal (B, C, and E).
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