Figure 1

Application of OpenEMMU to normal and cancer cell lines for analysis of DNA replication

(A) Diagram outlining the cell cycle phases, highlighting the S phase where thymidine analogs are incorporated during DNA replication.

(B) Graph depicting the annual number of PUBMED articles mentioning BrdU or EdU.

(C) Illustration of the EdU molecule and CuAAC reaction in cells that have incorporated EdU into nascent DNA.

(D) Flow cytometry results show the proportion of EdU-labeled cells after a 2-h pulse in indicated cell lines (n = 3).

(E) Relationship between EdU fluorescence intensity and the proportion of EdU+ cells at varying EdU concentrations in hiPSCs (n = 3).

(F) Comparison of EdU and DNA fluorescence intensity at different EdU concentrations from (E).

(G) Flow cytometry analysis of hiPSCs following a 1-h EdU pulse. The dotted lines indicate the gating strategy employed for both FACS and subsequent confocal microscopy of sorted cells (right panels) (n = 3).

(H) Confocal microscopy images of indicated EdU+ cancer cell lines after a 2-h EdU pulse, with α-Tubulin immunolabeling and DNA staining (n = 3). E: Early DNA replication (S-phase); M: Mid DNA replication; L: Late DNA replication.

(I) Cost per 0.5 mL reaction comparing OpenEMMU against two commercially available kit options (Invitrogen, kit 1; Vector Labs, kit 2).