Rabbit anti actin

Fat bodies of mosquitoes were dissected 12 h or 24 h post blood meal. Proteins of 10 mosquito fat bodies were extracted in 100 μl lysis buffer (50 mM Tris, pH 7.4; 1% IGEPAL 0.25% sodium deoxycholate; 150 mM NaCl; 1 mM EDTA; 1 mM phenylmethylsulfonyl fluoride; 1× protease inhibitor mixture; 1× phosphatase inhibitor mixture) [9 (link)]. Immunoblotting was performed using standard procedures. Antibodies used for TOR signaling were rabbit anti-phospho-S6K (Thr398) (1:1000) (Cell Signaling), rabbit anti-S6K (1:1000), and rabbit anti-actin (1:2000) (Sungenebiotech, China). Protein used for immunoblotting for p-Akt was extracted from fat bodies/ovaries, and midgut 12 hpi. The p-Akt was detected using a Phospho-Akt (Ser473) Antibody (1:200) (Cell Signaling) [51 (link)]. Immunoblotting for TEP1 was performed similarly, except that proteins from ten whole mosquitoes were extracted in cracking buffer (8 M urea, 2% SDS, 5% β-mercaptoethanol, 125 mM Tris-HCl) and 1:1000 anti-TEP1 rabbit polyclonal antibody was used. Intensity of the signals was quantified by ImageJ software [52 (link)].