Research Paper Volume 16, Issue 3 pp 2005—2025

Figure 6. Inhibitory effect of VitD on age- and oxidative stress-related accumulation of supernumerary centrosomes in midgut ISCs. (A) The guts from 10-day-old Myo^ts>GFP flies, 45-day-old Myo^ts>GFP flies, 10-day-old Myo^ts>GFP+Catⁿ¹ mutant flies, and 10-day-old Myo^ts>GFP+hr96Ri flies, without or with 100 nM VitD feeding for 1 week were stained with anti-PH3 (white), anti-γ-tubulin (red), anti-GFP (green), and DAPI (blue). The original magnification is 400×. (B) The number of PH3⁺ cells in whole guts from 10-day-old Myo^ts>GFP, 45-day-old Myo^ts>GFP, 10-day-old Myo^ts>GFP+Catⁿ¹, and Myo^ts>GFP+hr96Ri flies, with or without VitD feeding for 7 days was determined. Data (mean ± standard error) in 10-day-old Myo^ts>GFP, 45-day-old Myo^ts>GFP, 10-day-old Myo^ts>GFP+Catⁿ¹, and Myo^ts>GFP+hr96Ri flies without VitD feeding were collated from 165, 823, 1835, and 2484 mitotic cells of 25, 17, 16, and 22 guts, respectively. Data (mean ± standard error) in 10-day-old Myo^ts>GFP, 45-day-old Myo^ts>GFP, 10-day-old Myo^ts>GFP+Catⁿ¹, and Myo^ts>GFP+hr96Ri flies with VitD feeding were collated from 126, 489, 1235, and 1445 mitotic cells of 28, 18, 18, and 19 guts, respectively. n.s., no significant difference from the control (p > 0.05). (C) The frequency of supernumerary centrosomes (>2) per mitotic ISC in 10-day-old Myo^ts>GFP, 45-day-old Myo^ts>GFP, 10-day-old Myo^ts>GFP+Catⁿ¹, and Myo^ts>GFP+hr96Ri flies with or without VitD feeding for 7 days. The centrosome numbers in mitotic ISCs (PH3⁺ and GFP^- cells) in the midgut were determined. n.s., no significant difference from the control (p>0.05). (D) The frequency of mitotic ISCs with supernumerary centrosomes per gut in 10-day-old Myo^ts>GFP, 45-day-old Myo^ts>GFP, 10-day-old Myo^ts>GFP+Catⁿ¹, and 10-day-old Myo^ts>GFP+hr96Ri flies with or without VitD feeding for 1 week. Error bars represent standard error. P-values were calculated using Student’s t-test. n.s., no significant difference from the control (p>0.05).