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. 2022 Jun 9;17(7):1772–1785. doi: 10.1016/j.stemcr.2022.05.003

Figure 6.

Figure 6

Flow cytometric, immunostaining and qPCR analysis of cells in subtype-specific iPSC-CMs

(A) Immunostaining of cells differentiated using the standard protocol and purified using miR-208a-switch.

(B and E) Flow cytometric analysis of cells differentiated using the atrial (B) and nodal (E) protocols. (Left) The percentage of troponin T-positive cells and cells that reacted to miR-208a switch before purification. (right) Troponin T-positive cells were significantly increased (B, 95.4 ± 3.3%; and E, 95.2 ± 2.4%) after the final purification. The values were acquired from three biologically independent measurements.

(C and F) qPCR analysis of ventricular-related (MYL2 and IRX4), atrial-related genes (COUPTF-Ⅱ and MYL7) (C) and nodal-related genes (SHOX2 and ISL1) (F) in cells before purification, after purification with the conventional antibody-based method, and after purification with the miR-208a-switch. The values were acquired from three biologically independent measurements.

(D and G) Immunostaining of purified iPSC-CMs using miR-208a switch with the atrial (D) and nodal protocols (G). All scale bars, 100 μm. The values are denoted as means. All error bars represent the standard deviation. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001, by a paired t-test.