Journal: bioRxiv
Article Title: Replicable generation of rhesus macaque iPSCs for in vitro modeling of genetic frontotemporal dementia
doi: 10.64898/2026.03.17.712482
Figure Lengend Snippet: Characterization of RhiPSC line M6 clone 16.02. (A) Amplicon sequencing of MAPT R406W with mutant T allele indicated in the black box with a red arrow compared to reference genome (ref). (B) Phase contrast image of colonies grown on MEFs and (C) positive alkaline phosphatase staining; scale bar, 100 µm. (D) RT-PCR detection of endogenous pluripotent mRNAs POU5F1 , SOX2 , NANOG , KLF4 , LIN28 , and MYC . ACTB was used as a reference gene. (E) Amplification of EBNA and oriP in RhiPSCs at P18, no template control (NTC), and the rhesus embryonic stem cell line r420 (RhESC). Plasmid EM2K was used as a positive control (PC) for the detection of EBNA and oriP. (F) Positive immunofluorescence staining for pluripotency proteins SOX2 (green), OCT4 (red), and NANOG (far red), colocalized with DAPI (blue). Scale bar, 100 µm. (G) Flow cytometry-detected APC-A+ cells were 93.1% positive for SOX2, 98% positive for OCT4, and 93.3% positive for NANOG. (H) Short tandem repeat (STR) analysis confirmed (+) RhiPSC line identity. Mycoplasma (Myco.), Herpes B Virus (HBV), Simian Immunodeficiency Virus (SIV), Simian T-Lymphotropic Virus (STLV), and Simian Retrovirus (SRV) were not detected (-). (I) Normal 42,XX karyotype at P15. (J) H&E staining of a teratoma containing endoderm, mesoderm, and ectoderm. Scale bar, 100 µm.
Article Snippet: After 5 days, colonies were stained with a Vector Blue Alkaline Phosphatase Substrate kit (Vector Laboratories, cat. no. SK-5300) as described in .
Techniques: Amplification, Sequencing, Mutagenesis, Staining, Reverse Transcription Polymerase Chain Reaction, Control, Plasmid Preparation, Positive Control, Immunofluorescence, Flow Cytometry, Virus