Cell culture and osteogenic differentiation
Live-cell imaging
Western blot analysis
High-resolution SE-ADM
SE-ADM culture dish holder
Image processing
Calculation of particle diameters
Immunofluorescence staining of the cells and observation by optical microscopy
Immunolabeling of osteoblasts using antibody and 60-nm gold colloids
EDX spectrometric analysis
Raman microscopy
Transfection of CRISPR-Cas9 ribonucleoprotein complex
Single-cell sorting
Quantitative reverse transcription polymerase chain reaction
Genomic PCR and subcloning for sequencing analysis
Inhibitor treatment
The 80 μl of 2-week-culture supernatants from single cell–sorted 96-well plates were used to measure alkaline phosphatase activity with the Alkaline Phosphatase Assay Kit (ab83369; Colorimetric, Abcam, Cambridge, UK) according to the manufacturer’s instruction.