Figure 8.

Responses revealed were due to VK2 treatment. Loss of function of heix lead to mitochondrial defects represented by hypoxia. (A) Assessment of HIF protein expression by western blot analysis for conformation of hypoxia condition, distinct expression of HIF protein in heix mutants (heix ^k11403 /heix ¹) reared on control medium compared with a significant lesser expression in both control (Canton S) and VK2 treated groups. The difference in expression level quite significant. (A) Expression of mitochondrial regulator genes (Bcl2 family), a western blot including Bcl2, Bax, and P53, there is clear expression of Bax and P53 and fair expression of Bcl2 in heix mutants, all responded to treatment with VK2, and reversed expression was revealed. Response was significant with P53 protein, more significant with both Bax and Bcl2. (B) Treatment of heix mutant larvae with antioxidants N-Acetyl-L-cysteine (NAC) and (Mito-TEMPO) (MT) scavengers in three treatments group design, one with NAC, the other with MT and the third with both of them. heix mutant on control medium with distinct hyperplasia of lymph gland, control (Canton S) on control medium with normal lymph gland, heix mutant treated with MT scavenger showed lymph gland hyperplasia, heix mutant treated with NAC scavenger also showed lymph gland hyperplasia, heix mutant treated with MT + NAC scavengers revealed lymph gland hyperplasia, heix mutant treated with VK2 showed normal lymph gland. (C) Scavengers didn’t reduce the high lobulation of heix mutant lymph gland as VK2 achieved. (D) The percentage of black spots in heix mutant larvae didn’t affected by scavenger’s treatment as demonstrated with VK2 treatment which showed complete disappearance of them. Control media of control (Canton S) and heix mutant contain 50 mM Ethanol (EtOH), and treatment media contain 50 mM of each scavenger’s. Error bars SEM. A significant difference is compared to the control (Canton S) (ANOVA/Dunnett: *P < 0.05; **P < 0.01; ***P < 0.001).