Figure 3

Membrane ruffling effects induced by insulin, commercial and rice-produced rhIGF-I proteins on L6 myotubes. (A) Morphology of membrane ruffling effects induced by insulin and commercial rhIGF-I (200×). a) Normal L6 muscle cells. After adding b) 100 nM of insulin and c) 16 nM of commercial rhIGF-I, the actin filaments of L6 cells were reorganized and looked like human winkle. (B) Morphology of membrane ruffling effects induced by commercial and rice-produced rhIGF-I (SI) (200×). The ruffling effect was estimated by dividing each results photo into nine hundreds equal squares. Commercial rhIGF-I (4, 8 and 16 nM), SI seed crude protein extract (0.625 - 10 mg) were used to treat L6 cells. Wild type rice (12 mg) seed crude protein and protein extraction buffer (basal) were the negative controls. The average number of squares showing ruffling in any three photos was counted and the percentage of ruffling was calculated. The squares showing ruffling were marked in blue. a) Commercial rhIGF-I 4 nM; b) Commercial rhIGF-I 8 nM; c) Commercial rhIGF-I 16 nM; d) Wild type rice 12 mg; e) Protein extraction buffer, basal; f) SI 0.625 mg; g) SI 1.25 mg; h) SI 2.5 mg; i) SI 5 mg; J) SI 10 mg. (C) A graph showing membrane ruffling effects induced by commercial and rice-produced rhIGF-I (SI). The three columns on the left hand side represented the ruffling effect induced by 4, 8 and 16 nM of commercial rhIGF-I while the remaining indicated membrane ruffling caused by 0 - 10 mg of SI seed crude protein. Data are shown as means ± SD.