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. 2022 May 25;25(6):104459. doi: 10.1016/j.isci.2022.104459

Figure 3.

Figure 3

MASTL is highly expressed in pluripotent stem cells and supports stemness

(A) Expression of MASTL in the Amazonia database (http://amazonia.transcriptome.eu) (Gene expression Atlas for Embryonic SC, Probe 228468_at, U133P2) (mean + SEM, unpaired t-test).

(B) Western blotting of MASTL, OCT4, vimentin, and GAPDH in hiPSCs and differentiated fibroblasts.

(C) Expression of MASTL in differentiation to embryonic layers day 5 (Gifford et al., 2013), (mean + SEM, unpaired t-test).

(D) Western blotting of MASTL, NANOG, SOX2, and GAPDH in embryonic body (EB) differentiation in addition to phase-contrast images (scale bar 100 μm).

(E–H) Immunostaining of MASTL, NANOG, SSEA-5, and DAPI during reprogramming and quantification of the nuclear MASTL signal intensity in NANOG/SSEA-5 positive and negative cells (n = 3 biologically independent experiments, unpaired t-test, mean + SD). Scale bar, 10 μm.

(I) Western blotting of MASTL, β1-integrin, NANOG, SOX2, and GAPDH in original fibroblasts used for reprogramming and in hiPSCs.

(J) Western blotting of MASTL, OCT4, NANOG, and GAPDH in siControl-, siMASTL#6-, and siMASTL#7-treated hiPSCs for 48 h.

(K and L) Relative protein expression of OCT4 (K) or NANOG (L) to GAPDH, experimental setup shown in (J). (n = 3 biologically independent experiments, unpaired t-test, mean ± SD). See also Figure S3.