Dp controller manager software

In brief, 1 × 105 EAHY cells were seeded on the sterile coverslips in a 24-well plate in DMEM and 10% FBS. After 24 hours, they were incubated in DMSO or 20 μg/ml of 7-KC for further 24 hours. Medium was then aspirated, and cells were further rinsed with PBS and fixed in 4% paraformaldehyde for 20 min. EAHY cells were rinsed with PBS, permeabilized with 2% Triton X-100, treated by 0.3% v/v H₂O₂ for 20 minutes. After further washed by PBS, cells were blocked in 5% bovine serum albumin (BSA) for 1 hr and then incubated in primary antibodies (p-ATM, p-ATR, p-Chk1, p-Chk2, p-p53, p-Akt) (1:1000, v/v) at room temperature for overnight. Following PBS wash, cells were incubated in respective secondary antibody (FITC- or TRITC-conjugated) in the dark for 1 hr and counterstained with 4',6-diamidino-2-phenylindole (DAPI, 1:1000) for 30 min. Finally the cell coverslips were mounted and observed/photographed by an Olympus IX71 inverted microscope and DP Controller/Manager software (Olympus Corporation, Tokyo, Japan) [36 (link)].

In brief, 4 × 10⁴ human dental pulp cells were seeded on the sterile coverslips in a 24-well plate in DMEM and 10% FBS. After 24 hours, they were exposed to different concentrations of CQ (0.25, 0.5, 1, 2, 3 mM) for further 24 hours. Medium was decanted, and cells were washed with PBS and fixed in 4% paraformaldehyde for 20 minutes. Cells were washed with PBS, permeabilized with 2% Triton X-100, incubated in 0.3% v/v H₂O₂ for 20 minutes. After washed with PBS, cells were blocked in 5% bovine serum albumin (BSA) for 1 hour and then incubated in primary antibodies (p-ATM, p-Chk2, p-p53 and GADD) (1:1000, v/v) at room temperature overnight. After PBS wash, cells were incubated in corresponding secondary antibody (FITC- or TRITC-conjugated) in the dark for 1 hour and counterstained with DAPI (1:1000) for 30 minutes. Finally the samples were mounted and observed/photographed under Olympus IX71 inverted microscope and DP Controller/Manager software (Olympus Corporation).

Briefly, 1 x 10⁵ of MG-63 cells were seeded on the sterile coverslips in a 24-well plate in DMEM and 10% FBS. After 24 h, they were subjected to different concentrations of butyrate (0–16 mM) for additional 24 h. Medium was decanted, and cells were rinsed with PBS and fixed for 20 min in 4% paraformaldehyde. Cells were washed by PBS, membrane penetration with 2% Triton X-100, incubated for 20 min in 0.3% v/v H₂O₂. After washed with PBS, cells were blocked in 5% bovine serum albumin (BSA) for 1 h and then incubated in primary antibodies (against type I collagen) at room temperature overnight. After washing by PBS, cells were incubated in corresponding secondary antibody in the dark for 1 h and counterstained for 30 min with 4',6-diamidino-2-phenylindole (DAPI, 1:1000) [17 (link)]. Finally the samples were mounted and observed/photographed by an Olympus IX71 inverted microscope and DP Controller/Manager software (Olympus Corporation).