Fig. 4: In vitro and in vivo bioactivity of FHI after prolonged exposure to higher temperature.
From: Synthesis of a highly thermostable insulin by phenylalanine conjugation at B29 Lysine
a Representative immunoblots showing the relative levels of P-AKT (Ser473) in the HEK293T cell lines that are untreated (UT) or treated for 30 min with 0.5 µM of HI or FHI incubated at 65 °C for different time points as indicated. γ-tubulin served as the loading control; b Plot showing the change in the activity of P-AKT in the incubated HI or FHI-treated HEK293T cells as compared to the untreated (UT) control at the indicated time points. (N = 3, Error bars represent SE of mean); c Blood glucose level in STZ-treated C57BL/6 J wild-type mice after subcutaneous injection (1 µg/100 µL per 30 gm of mice) of heated human insulin and the modified insulin FHI. Heating was carried out in 0.1 N HCl water (pH 1.6) containing 25 mM NaCl at 65 °C for 12 h and 48 h for HI and FHI respectively. Data are represented as the mean of each group (N = 3) for each time point. Error bars represent the SE of the mean.
