Fig. 2: THEMIS2 was able to promote cancer stemness properties in TNBC cells. | Oncogene

Fig. 2: THEMIS2 was able to promote cancer stemness properties in TNBC cells.

From: Novel function of THEMIS2 in the enhancement of cancer stemness and chemoresistance by releasing PTP1B from MET

Fig. 2

A The protein expression levels of CSC markers in THEMIS2-overexpressing MDA-MB-231 and Hs578T cells, relative to those of their parental cells, were analyzed through immunoblotting. Actin was used as an internal control. B The sphere-forming abilities of three stable THEMIS2-overexpressing TNBC lines (MDA-MB-231, Hs578T, and BT549) relative to those of their parental cells. The histograms represent means ± SEs from three independent experiments (**P < 0.01; ***P < 0.001). C The colony-forming abilities of three stable THEMIS2-overexpressing TNBC lines (Hs578T, BT549, and MDA-MB-231-IV2 (IV2)) in soft agar, relative to those of their parental cells, were analyzed through a soft agar colony formation assay. The histograms represent means ± SEs from three independent experiments (**P < 0.01). D The invasion abilities of three stable THEMIS2-overexpressing TNBC lines (Hs578T, MDA-MB-231, and BT549), relative to those of their parental cells, were analyzed using a chemotactic cell invasion assay. The histograms represent means ± SEs from three independent experiments (**P < 0.01). E 5 × 102 stable MDA-MB-231-pCMV6-THEMIS2 cells or stable pCMV6-control cells were orthotopically implanted into the mammary fat pads of 10 CB17-SCID mice (n = 10 for each group). Representative images of tumor growth and tumor latency are shown. F Left: THEMIS2 protein expression levels in stable THEMIS2-overexpressing MDA-MB-231 cells, as determined by immunoblotting analysis. Actin served as an internal control. The BLI image scatter plots represent means ± SEs from three independent experiments (*P < 0.05). Right: representative images of pulmonary metastasis induced by 5 × 105 MDA-MB-231 cells with or without THEMIS2 overexpression in the CB17-SCID mice, 21 days after implantation (n = 3 for each group). Tumor growth was monitored through bioluminescent imaging. The actin bands in A served as the representative loading control of multiple immunoblots from the same batch of cell lysates. They served as the internal controls for comparing the intensity of the proteins bands stained by a given antibody.

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