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The fluorescence intensity from APF that has been oxidized by H2O2 is pH dependent (Figure 4).
APF was added to MPO alone or in the presence of SIN-1/peroxynitrite at 37 °C and the formation of fluorescein from APF was measured every 30 s for indicated time periods.
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Briefly, retinae were dissected from wandering third instar larvae or from 42H apf pupae in HCO3 buffer (25 mM NaHCO3, 115 mM NaCl, 5 mM KCl, 10 mM glucose, 1 mM MgSO4, 1 mM KHPO4, 2 mM CaCl2) and mounted in MatTek glass-bottom dishes.
Gene expression was assayed at five time points in animals collected every 24 hours, ranging from 0 hours after puparium formation (APF; 0 hour APF is the white pre-pupal stage) to 96 hour APF (pharate adults).
Wg is required from 15 20 hr APF for bristle formation and from 18 28 hr APF for tergite differentiation and pigmentation.
(B ) Still images taken from time-lapse videos at 24, 33 and 42 hr APF from control, miR-965 mutant and rescued mutant to illustrate expansion of the histoblast nests to replace LECs.
3'- p-aminophenyl) fluorescein (APF) from Invitrogen™ was used for the detection of hROS.
Our simulation results suggest that, despite the reduction in the number of particles from 8000 to 5000, the APF tracker still outperforms the RS tracker, showing similar RMS error performance with a slightly lower divergence rate.
All pupae were staged from 0 h APF, defined as stiff white pre-pupae (approximately 120 h AED ± 20 min at 25°C).
We performed TUNEL assay with wild-type and grim A6C homozygous CNSs dissected from 6 8 hours APF, a period that we previously observed most abundant TUNEL signals (Lee et al., 2011).
Interestingly, Aret is required for both splice suppression of a tubular muscle-specific exon and inclusion of a fibrillar muscle-specific exon from 30 h APF onwards (Supplementary Fig S5K).
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