Fig. 2

Preliminary tests confirming the accuracy of single-cell RNA-snMIFxC. (A) Sorted HeLa and human deciduous tooth-derived dental pulp cells (HDDPCs). There are ~ 10 HeLa cells with high alkaline phosphatase (ALP) activity (a) and those with less ALP activity (b). Similarly, there are ~ 10 HDDPCs with high ALP activity (c) and those with less ALP activity (d). Bar = 10 μm. (B) Reverse transcription-polymerase chain reaction (RT-PCR) using whole transcriptome amplification (WTA) products derived from HDDPC-ALP( +), HDDPC-ALP(-), HeLa cells-ALP( +) and HeLa cells-ALP(-) to amplify transcripts of glyceraldehyde 3-phosphate dehydrogenase (GAPDH). M, 100 bp ladder; -RT, PCR, performed in the absence of reverse transcription products; HDDPC PC, RT-PCR products using cDNAs derived from unselected HDDPCs; iPSC PC, RT-PCR products using cDNAs derived from unselected HDDPC-derived induced pluripotent cells (iPSCs). (C) RT-PCR to check the expression of ALP, octamer-binding transcription factor 3/4 (OCT3/4) and sex determining region Y-box 2 (SOX2) in HDDPC-ALP( +) and HDDPC-ALP(-) using appropriate primer sets. In the right panel, comparison of the expression levels of each transcript between HDDPC-ALP( +) and HDDPC-ALP(-), after normalization is shown. The intensity of each band in the left panel was determined using the ImageJ software. The level of each transcript was normalized to that of GAPDH mRNA, and the results obtained are represented as a graph. M, 100 bp ladder; -RT, PCR performed in the absence of reverse transcription products. HDDPC PC, RT-PCR products using cDNAs derived from unselected HDDPCs; iPSC PC, RT-PCR products using cDNAs derived from unselected HDDPC-derived iPSCs