Fig. 2 | Oncogene

Fig. 2

From: Differential regulation of the sphere formation and maintenance of cancer-initiating cells of malignant mesothelioma via CD44 and ALK4 signaling pathways

Fig. 2

HA synthesis is upregulated in MM spheroids. a MSTO-211H cells were cultured for 4 days under conventional adherent or sphere-forming conditions. Total RNA was isolated from the adherent cells or spheroids, and the expression of HAS1, HAS2, and HAS3 was determined by RT-PCR. b ALDH activity in MSTO-211H spheroids was determined by ALDEFLUOR assay, and the cell-surface expression of CD44 in the ALDHbright (red) or ALDHdim (blue) cell populations was analyzed by flow cytometry. c MSTO-211H spheroids were fixed with PFA and stained for hyaluronan-binding protein (HABP), anti-CD44, and DAPI (upper). To verify the specificity of the HABP staining, hyluronidase (HAase) treatment was performed before staining (lower). d MSTO-211H spheroids were treated with HAase and incubated with FL-HA in the presence of control or neutralizing anti-CD44. e Ki-67+ cells in MSTO-211H spheroids were analyzed by flow cytometry (upper) and immunohistochemistry (lower)

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