Supplementary material

Roles of catalase and glutathione peroxidase in the tolerance of a

pulmonate gastropod to anoxia and reoxygenation

Alexis F. Welker1,2, Daniel C. Moreira1 and Marcelo Hermes-Lima1,*

1Laboratório de Radicais Livres, Departamento de Biologia Celular, Universidade de Brasília,

Brasília, DF, 70910-900, Brazil.

2Faculdade da Ceilândia, Universidade de Brasília, Ceilândia, DF, 72220-140, Brazil.

*Corresponding author:

Dr. Marcelo Hermes-Lima

Laboratório de Radicais Livres, Departamento de Biologia Celular, Universidade de Brasília, Brasília,

DF, 70910-900, Brazil.

Phone: +55-61-3107-3103

Email: hermes@unb.br , hermes.unb@gmail.com and m_hermes_lima@yahoo.com.br

Fig. S1 Effects of ATZ injection and reoxygenation on total glutathione peroxidase (total-GPX) activity (mU/mg of protein),

glutathione transferase (GST) activity (U/mg of protein) and total glutathione equivalents (GSH-eq) levels (nmol/g of wet

weight) in hepatopancreas (a, b, c) and foot muscle (d, e, f) of Helix aspersa snails. Activity of total GPX (a, d) and GST (b,

e) and GSH-eq levels (c, f) in hepatopancreas and foot muscle from saline- or ATZ-injected snails exposed to anoxia and at

different reoxygenation times “zero” (5 h anoxia), 0.25 h, 0.5 h, 1 h, 2 h and 12 h from saline-injected and ATZ-injected snails.

There were no statistically significant alterations. (n=5–6)

Fig. S2 Effects of anoxia and ATZ injection (a,c) and of reoxygenation (b,d) with catalase inhibition

or not on reduced glutathione (GSH) levels (nmol/g of wet weight) of Helix aspersa snails. GSH

concentration in hepatopancreas (a) and foot muscle (c) from control (Ctrl, no injection), saline- (Sal,

animals injected with 0.9% (w/v) NaCl) or ATZ-injected (ATZ, animals injected with ATZ at 1

mg/gww solubilized in 0.9% NaCl) snails maintained in normoxia or exposed to anoxia. GSH

concentration in hepatopancreas (b) and foot muscle (d) at different reoxygenation times “zero” (5 h

anoxia), 0.25 h, 0.5 h, 1 h, 2 h and 12 h from saline-injected and ATZ-injected snails. There were no

statistically significant alterations. (n=5–6)

Fig. S3 Effects of anoxia and ATZ injection (a,c) and of reoxygenation (b,d) with catalase inhibition

or not on disulfide glutathione (GSSG) levels (nmol/g of wet weight) of Helix aspersa snails. GSSG

concentration in hepatopancreas (a) and foot muscle (c) from control (Ctrl, no injection), saline- (Sal,

animals injected with 0.9% (w/v) NaCl) or ATZ-injected (ATZ, animals injected with ATZ at 1

mg/gww solubilized in 0.9% NaCl) snails maintained in normoxia or exposed to anoxia. GSSG

concentration in hepatopancreas (b) and foot muscle (d) at different reoxygenation times “zero” (5 h

anoxia), 0.25 h, 0.5 h, 1 h, 2 h and 12 h from saline-injected and ATZ-injected snails. There were no

statistically significant alterations. (n=5–6)