For clonality assays, cells were plated on day 0 on clonal dilutions and stained on day 7.
Then, the cells were differentiated as described under 'Cell culture and differentiation', with addition of doxycycline in the medium until day 2. The cells were fixed and stained at day 7 of differentiation.
2) Similar procedures were used for the comparison of IPC groups subject to SOD or vehicle treatment (3 groups), and for the comparison of staining at day 3, day 6 and control.
Cultures were stained on day 14 post-induction with Oil-Red-O, as previously described [27,32].
To assess the reprogramming efficiency, plates were stained on day 14 for alkaline phosphatase (AP) activity (Vector Red Alkaline Phosphate substrate kit).
Intervention: Exposure to PSWD, twice daily, on days 2, 3, and 4. Main Outcome Measure: After crystal violet staining (day 5), optical density (cell number) was determined spectrophotometrically.
Furthermore, a greater number of B16F10 cells were detected in the lung tissues of RNF13-KO mice by frozen section at 6 h and 24 h or by H&E staining at day 2 and day 4.
Although there were still iodine molecules in the solution on day 5, the staining effect appeared to be stagnant in the latter stages of the first-round staining (day 5-day 10), and also for the rest of first-round staining (~ day 23).
A small portion (from none to < 5%, varied among donors) of Day 0 cells showed weak p53 staining (Day 0, Fig. 2B, red), while a bigger portion of cells (from 10%to30%0%, varied among donors) were positive for p16 (Day 0, Fig. 2C, red), suggesting that these cells might be undergoing senescence in vivo.
The painting titled "Click" is stained with Day-Glo orange of shifting intensities; the yellow-brown in "Daddy, Daddy, Daddy" seems to be applied in patches like gold on a Japanese screen.
Then, cells were washed with Dulbecco's PBS (Amimed, Bioconcept, Switzerland), fixed with 4% PFA for 15 minutes and permeabilized with 1% Triton X-100 in PBS for 15 minutes at RT and stained with Mon-152 (1∶100, Alexis Corporation).
