Journal: Oncology reports

Article Title: Hsp90 inhibitor 17‑AAG inhibits stem cell‑like properties and chemoresistance in osteosarcoma cells via the Hedgehog signaling pathway.

doi: 10.3892/or.2020.7597

Figure Lengend Snippet: Figure 2. Hsp90 is positively associated with the expression of stem cell‑related genes and CSC markers in osteosarcoma sarcosphere cells. (A) mRNA expres- sion levels of Hsp90 and the stem cell markers Sox 2, Oct‑4 and Nanog in sarcosphere and monolayer cells derived from MG‑63 and Saos‑2 cells. (B) Double staining for Hsp90 and Sox 2 using immunofluorescence in sarcosphere cells derived from MG‑63 and Saos‑2 cells. Scale bar, 50 µm. (C) Analysis of Hsp90, CD133, ALDH1 and CD271 expression in sarcosphere and monolayer cells derived from MG‑63 and Saos‑2 cells Hsp, heat shock protein; CSC, cancer stem cell; Sox 2, sex determining region Y‑box 2; Oct‑4, octamer‑binding transcription factor 4; ALDH1, aldehyde dehydrogenase 1. **P<0.01, ***P<0.001.

Article Snippet: Cells were detached into single-cell suspensions using trypsin-EDTA, followed by staining of 1x106 cells in 500 μl PBS/0.5% BSA (Sigma-Aldrich; Merck KGaA) with fluorescence‐labeled primary antibodies (1‐5 μl), including CD133-PE (1:10; cat. no. 130-112-195; Miltenyi Biotec GmbH), AldeflourTM kit (cat. no. 01700; Stemcell Technologies, Inc.) and CD271‐fluorescein isothiocyanate (1:400; cat. no. 746728; BD Biosciences) at 4 ̊C for 60 min. After washing, labeled cells were analyzed and sorted immediately using a BD FACS AriaIII system (BD Biosciences).

Techniques: Expressing, Derivative Assay, Double Staining, Immunofluorescence