Figures & data
Note: Original magnification: 100× and 400×.
Abbreviations: ESCC, esophageal squamous cell carcinoma; IHC, immunohisto-chemistry.
Abbreviations: ESCC, esophageal squamous cell carcinoma; IHC, immunohisto-chemistry.
Notes: The ESCC cell lines KYSE510 and KYSE150 were transiently transfected with two independent KIAA1522-specific siRNA (si-A and si-C) or negative control siRNA (siCtrl) for 48 h. (A) The expression levels of KIAA1522 protein were detected by immunoblotting. The arrow indicates the specific band of KIAA1522 protein located around 170 kDa. Relative ratios of absorbance for KIAA1522 to GAPDH were plotted. The data are presented as the mean ± SEM. ***P<0.001. (B) Cell proliferation was determined by CCK-8 assay. Data are mean ± SEM. ***P<0.001. (C) Colony formation assay. Representative results are shown, and the colony number was plotted. The data are presented as the mean ± SEM from three separate experiments. ***P<0.001.
Abbreviations: CCK-8, cell counting kit-8; Ctrl, control; ESCC, esophageal squamous cell carcinoma; GAPDH, glyceral dehyde 3-phosphate dehydrogenase; P, parental; SEM, standard error of the mean; si, siRNA; si-A, siKIAA-A; si-c, siKIAA-C.
Abbreviations: CCK-8, cell counting kit-8; Ctrl, control; ESCC, esophageal squamous cell carcinoma; GAPDH, glyceral dehyde 3-phosphate dehydrogenase; P, parental; SEM, standard error of the mean; si, siRNA; si-A, siKIAA-A; si-c, siKIAA-C.
Notes: (A) KYSE510 and KYSE150 cells were cultured in suspension for the indicated times, and the KIAA1522 protein levels were detected by Western blotting. Relative ratios of absorbance for KIAA1522 to GAPDH were plotted. The data are presented as the mean ± SEM. **P<0.01. The arrows in images A and B indicate the position of KIAA1522 specific band. (B–D) KYSE150 and KYSE510 cells were transiently transfected with KIAA1522 siRNA for 48 h and then cultured on polyHEMA-coated dishes for 20 h. (B) KIAA1522 expression was determined by Western blot. (C) Anoikis was measured by Annexin V-FITC/PI staining and flow cytometry analysis. Representative results are shown (upper panel), and the percentages of apoptotic cells were plotted (lower panel). The data are presented as the mean ± SEM. **P<0.01, ***P<0.001.
Abbreviations: Ctrl, control; ESCC, esophageal squamous cell carcinoma; FITC, fluorescein isothiocyanate; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; NC, non-silencing; P, parental; PI, propidium iodide; polyHEMA, polyhydroxylethylmethacrylate; si, siRNA; si-A, siKIAA-A; si-B, siKIAA-B; si-c, siKIAA-C.
Abbreviations: Ctrl, control; ESCC, esophageal squamous cell carcinoma; FITC, fluorescein isothiocyanate; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; NC, non-silencing; P, parental; PI, propidium iodide; polyHEMA, polyhydroxylethylmethacrylate; si, siRNA; si-A, siKIAA-A; si-B, siKIAA-B; si-c, siKIAA-C.
Notes: (A) KYSE510 and KYSE150 cells transfected with shKIAA or shCtrl were cultured on polyHEMA-coated dishes for 20 h and subjected to Annexin V-FITC/PI staining followed by flow cytometry analysis. Representative results are shown, and degrees of apoptosis were plotted. The data are presented as mean ± SEM. *P<0.05. (B–D) Nude mice were injected with 2×106 KYSE150-shCtrl or KYSE150-shKIAA stable cell clones via the tail vein. After 4 weeks, the mice were sacrificed. (B) The lungs were subjected to Bouin’s fixation and photographed. Scale bar =1 cm. (C) Visible lung metastases nudes were counted and plotted. (D) The sections of the lungs were stained with H&E. Scale bar =200 μm.
Abbreviations: Ctrl, control; NOD/SCID, nonobese diabetic/severe combined immunodeficiency; polyHEMA, polyhydroxylethylmethacrylate; sh, shRNA.
Abbreviations: Ctrl, control; NOD/SCID, nonobese diabetic/severe combined immunodeficiency; polyHEMA, polyhydroxylethylmethacrylate; sh, shRNA.
Notes: (A) The ESCC cell lines KYSE510 and KYSE150 were transiently transfected with KIAA1522-specific siRNA or control non-silencing siRNA for 48 h. (B) Transiently transfected cells were cultured on polyHEMA-coated dishes for 20 h, and the cell lysates were immunoblotted for the indicated proteins. The arrows indicate the position of KIAA1522 specific band. Relative ratios of absorbance for KIAA1522 to GAPDH, pERK to total ERK were plotted. The data are presented as the mean ± SEM. **P<0.01, ***P<0.001.
Abbreviations: Ctrl, control; ERK, extracellular signal-regulated kinase; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; NC, non-silencing; P, parental; pERK, phosphorylated ERK; polyHEMA, polyhydroxylethylmethacrylate; si, siRNA; si-A, siKIAA-A; si-c, siKIAA-C.
Abbreviations: Ctrl, control; ERK, extracellular signal-regulated kinase; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; NC, non-silencing; P, parental; pERK, phosphorylated ERK; polyHEMA, polyhydroxylethylmethacrylate; si, siRNA; si-A, siKIAA-A; si-c, siKIAA-C.