Fig. 4: MST4 mediates the oncogenic effect of LIMK2 in ESCC. | Oncogene

Fig. 4: MST4 mediates the oncogenic effect of LIMK2 in ESCC.

From: LIMK2 promotes centrosome clustering and cancer progression by activating MST4-mediated phosphorylation of NPM1

Fig. 4

A Western blot of MST4 and GAPDH (loading control) of cells treated with MST4 shRNA. B MTT assay was measured for indicated cells (n = 3, independent experiments). C Resulting cells were subjected to soft agar assays and the relative colony numbers were quantified (n = 3, independent experiments). D Colony number were conducted in KYSE150 and KYSE450 cells treated with MST4 shRNA. (n = 3, independent experiments). Colony number was analyzed using Image J software, with cell size larger than 20 defined as valid clones. Colony numbers are as follows: KYSE150 (Mock: 55.7 ± 2.9, shMST4#2: 15.3 ± 2.1, shMST4#3: 5.3 ± 2.5), and KYSE450 (Mock: 66.3 ± 11.9, shMST4#2: 4.7 ± 2.1, shMST4#3: 5.3 ± 2.3). E Western blot assay of MST4 and Flag in indicated cells. GAPDH served as loading control. F MTT assay was measured for MST4-overexpressed cells (n = 3, independent experiments). G Colony formation assay in MST4-overexpressed cells (n = 3, independent experiments). Colony number was analyzed using Image J software, with cell size larger than 20 defined as valid clones. Colony numbers are as follows: KYSE30 (Vector: 10.3 ± 1.2, OE-MST4: 19.0 ± 2.6) and KYSE410 (Vector: 74.7 ± 6.7, OE-MST4: 101.7 ± 4.6). H Cells with LIMK2 overexpression and MST4 knock down was established, and the expression of Flag-LIMK2 and MST4 was detected by Western blot. GAPDH served as loading control. I Colony formation was performed for the indicated cells (n = 3, independent experiments). Colony number was analyzed using Image J software, with cell size larger than 20 defined as valid clones. Colony numbers are as follows: KYSE150 (Vector: 18.7 ± 3.2, Flag-LIMK2: 31.3 ± 4.7, shMST4: 10.3 ± 1.5, shMST4-OE-LIMK2: 10.0 ± 1.0) and KYSE450 (Vector: 17.7 ± 3.1, Flag-LIMK2: 33.3 ± 4.5, shMST4: 3.7 ± 1.5, shMST4-OE-LIMK2: 4.0 ± 1.0). J MTT assay was measured for indicated cells (n = 3, independent experiments). All data were shown as means ± SD and were analyzed with the two-tailed Student’s t test.*p < 0.05, **p < 0.01, ***p < 0.001.

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