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

Figure 5.

Figure 5

MASTL supports TGFBR2 levels

(A) Relative TGFBR2 mRNA expression in siControl-, siMASTL#6-, and siMASTL#7-treated MDA-MB-231 cells after 48 h of silencing (n = 5 biologically independent experiments, unpaired t-test, mean ± SD).

(B) Western blotting of MASTL, TGFBR2, and GAPDH in siControl-, siMASTL#6-, and siMASTL#7-treated MDA-MB-231 cells after 48 h.

(C) TGFBR2 protein expression relative to GAPDH in siControl-, siMASTL#6-, and siMASTL#7-treated MDA-MB-231 cells after 48 h (n = 3 biologically independent experiments, unpaired t-test, mean ± SD).

(D) Western blotting of MASTL, TGFBR2, and GAPDH in siControl-, siMASTL#6-, siMASTL#7-, siTGFBR2#5-, and siTGFBR2#6-treated MDA-MB-231 cells after 48 h. Cells were stimulated with TGF-β (20 ng/mL) for 45 min immediately before collection.

(E) TGFBR2 expression in breast cancer cell lines with high or low MASTL expression based on CCLE data (unpaired t-test, mean ± SEM).

(F) Western blotting of MASTL, TGFBR2, and GAPDH in EGFP (control), EGFP MASTL wild-type (WT), and EGFP MASTL kinase-dead (G44S) overexpressing MCF10A cells 24 h after transfection.

(G) TGFBR2 protein expression relative to GAPDH in EGFP (control), EGFP MASTL wild-type (WT), and EGFP MASTL kinase-dead (G44S) overexpressing MCF10A cells after 24 h (n = 3 biologically independent experiments, one-sample t-test, mean ± SD).

(H) Western blotting of MASTL, TGFBR2, and GAPDH in DMSO- and GKI-1 (20 μM)-treated MDA-MB-231 cells for 48 h.

(I) TGFBR2 protein expression relative to GAPDH in DMSO- and GKI-1-treated MDA-MB-231 cells after 48 h (n = 3 biologically independent experiments, one-sample t-test, mean ± SD). See also Figure S5.