Journal: Biomedicines
Article Title: P53 Deficiency Accelerate Esophageal Epithelium Intestinal Metaplasia Malignancy
doi: 10.3390/biomedicines11030882
Figure Lengend Snippet: P53 deficiency affects bile acid-mediated cell proliferation repression and intestinal metaplasia. ( A ) HET1A cells with P53 knockdown by shRNA, Western Blot assay of P53 expression level. ( B ) The proliferation of HET1A cells with or without P53 knockdown after 30 days of continuous exposure to ABS or BS. ( C ) Representative immunofluorescence of Ki67 images of HET1A cells with 400 μmol/L ABS or BS treatment. ( D ) Representative immunofluorescence of Ki67 images of P53 knockdown HET1A cells with 400 μmol/L ABS or BS treatment. ( E ) Quantification of Ki67 expression levels in HET1A cells with or without P53 knockdown stimulated by ABS and BS. ( F ) After continuous exposure to ABS for denoted time, the expression levels of squamous epithelium marker CK13 , columnar epithelium marker CK8 and intestinal metaplasia marker CDX2 . ( G ) After continuous exposure to BS for denoted time, the expression levels of CK13 , CK8 , and CDX2 . +: Treated accordingly; −: Not treated accordingly. Data were statistically analyzed using the student t -test. * p < 0.05, ** p < 0.01, ns: none significance.
Article Snippet: Next, the PVDF Blotting Membrane was blocked with 5% fat-free dry milk at room temperature for 2 h and incubated with the primary antibodies against CDX2 (1:1000; Proteintech; Cat. No. 60243-1-Ig), CK8 (1:1000; Proteintech; Cat. No. 17514-1-AP), CK13 (1:1000; Proteintech; Cat. No. 10164-2-AP), ICAM1 (1:1000; Proteintech; Cat. No. 10831-1AP), OCLN (1:1000; ABCAM; Cat. No. ab31721), CLDN2 (1:1000; ABCAM; Cat. No. ab53032), P53 (1:1000; Proteintech; Cat. No. 60283-2-Ig), P21 (1:1000; Proteintech; Cat. No. 10355-1-AP), JAK1 (1:1000; Proteintech; Cat. No. 66466-1-Ig), STAT3 (1:1000; Proteintech; Cat. No. 10253-2-AP), VEGFR (1:1000; Proteintech; Cat. No. 13687-1-AP), P-AKT (1:1000; Proteintech; Cat. No. 28731-1-AP), AKT (1:1000; Proteintech; Cat. No. 10176-2-AP), GAPDH (1:10,000; Proteintech; Cat. No. 60004-1-Ig), α-Tubulin (1:10,000; Proteintech; Cat. No. 66031-1-Ig) at 4 °C for overnight.
Techniques: shRNA, Western Blot, Expressing, Immunofluorescence, Marker
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