Dp74 ccd camera

Cells re-suspended in culture medium were seeded in 12-well plates (3 × 10⁵ cells per well). To inhibit p53, the cells were pretreated with pifithrin-α (20 μM) (#S5791 from Selleck Chemicals, Houston, TX, United States). After various treatments as indicated, the cells were pulse labeled with BrdU (10 μM) (from MedChemExpress, NJ, United States) for 1.5 h. BrdU incorporation was detected using immunofluorescence with a monoclonal BrdU antibody (#A20790, from ABclonal, Woburn, MA, United States) followed by Alexa Fluor 488-conjugated secondary antibody. The images were taken using a fluorescence microscope equipped with a DP74 CCD camera (Olympus, Tokyo, Japan).

The KC1-P2 and KC15-24 strains were observed using an Axio Imager.A2 microscope (Carl Zeiss, Berlin, Germany) equipped with an Olympus DP71 or DP74 CCD camera (Olympus, Tokyo, Japan). Endosymbiont-bearing KC1-P2 was observed using fluorescence microscopy. Fixed cells were stained with 4,6-diamidino-2-phenylindole (DAPI) in the dark and mounted with SlowFade DIAMOND (Invitrogen, Carlsbad, CA). Specimens were then observed under a Leica DMRD microscope (Leica, Wetzlar, Germany) equipped with an Olympus DP73 CCD camera (Olympus, Tokyo, Japan). For transmission electron microscopy (TEM) observations of the C. parkeae stage of B. bigelowii and its endosymbiont, the MK90-06 strain was used. Detailed methods for sample treatment and observation are described in Kawachi et al. (1991) (link) (Supplementary Material).

Root tips of 10-d-old seedlings grown on ½MS plate were treated with H₂O or 1 μM Pep1 on plates for 24 hours. H₂O₂ in root tips was detected with 2′,7′-dichlorofluorescein diacetate (H₂DCF-DA) (Sigma-Aldrich, St. Louis, MO, USA) staining. In brief, the seedlings were incubated in 25 μM H₂DCF-DA solution for 10 minutes in darkness. After three washes with water, seedlings were photographed under fluorescence microscopy (BX53, Olympus, Tokyo, Japan) equipped with DP74 CCD camera (excitation, 460 nm; emission, 520 nm).