Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Interleukin-23 receptor signaling mediates cancer dormancy and radioresistance in human esophageal squamous carcinoma cells via the Wnt/Notch pathway

doi: 10.1007/s00109-018-1724-8

Figure Lengend Snippet: The correlation between IL-23/IL-23R and the trans-differentiation of ESCCs. a Typical immunofluorescence images of the distribution of Oct-4A + cells (red) and IL-23R + ESCCs (green) gathering with the intensity of IL-23 (IHC). Left panel, × 200 magnification. Middle and right panels are magnifications of the area marked by dashed lines. b Western blotting: the expression of CD133 in ESCC cells (TE-1, ECA 109, KYSE 150, and TE-10) and Het-1A cells with or without IL-23 treatment (50 ng/mL, 24 h). The results were normalized to β-actin as a control and densitometric analysis of bands was performed with Alpha View. T, TE-1 cells; E, ECA 109 cells; H, Het-1A cells. c The percentage of CD133 + cells in ESCCs and Het-1A cells before and after sorting. Flow cytometry and fluorescent cell sorting were performed using anti-CD133 fluorescent-labeled antibody. The representative experiment results were compared with that untreated groups. d The number of protogenetic IL-23R + cells. Flow cytometry was performed using anti-IL-23R (FITC) in ESCCs and Het-1A cells. The experiment was repeated twice and representative data shown. e The variants of CD133 + cells between IL-23R − /IL-23R + CD133 − ESCCs and Het-1A cells cultured with IL-23 (50 ng/mL, 24 h). f CD133 − IL-23R + ESCCs and Het-1A cells were pretreated with IL-23 (50 ng/mL) for 24 h, the expression levels of CD133 were detected by Western blotting at 0, 24, 48, and 72 h after removing IL-23. g The relative mRNA expression levels of stemness genes (c-myc and Oct-4A) were measured by RT-PCR in CD133 − IL-23R + ESCCs and Het-1A cells cultured with IL-23 (50 ng/mL) for 24 h. The GAPDH was used as the loading control. TE-1, ECA 109 cells, and Het-1A cells were treated with IL-23 for 48 h or not, and the protein expression of Oct-4A and c-myc was determined by Western blot analysis. β-Actin was used as a loading control. The data are presented as the mean ± SD from at least three independent experiments. ** p < 0.01

Article Snippet: Immunohistochemical (IHC) and immunofluorescence staining, scoring and statistics were performed as previously described [ ], and primary antibodies for Oct-4A, CD133, IL-23, IL-23R, CD68, and HLA-DR were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Immunofluorescence, Western Blot, Expressing, Control, Flow Cytometry, FACS, Labeling, Cell Culture, Reverse Transcription Polymerase Chain Reaction