Figure 3
5-ASA promotes wound healing. (A,B) In vitro scratch assay on HCEC-1CT monolayer. Closure of scratch (wound healing) was observed under inverted microscope and pictures were taken at different time points indicated. Image magnification 100x. Pictures are representative of three independent experiments. Quantification was performed on two images. ANOVA with Tukey’s multiple comparisons test was performed (C) MTT based cell proliferation assay (48 h) in the presence or absence of 5-ASA (5 mM), AZTP (10 μM) or in combination was performed (n = 4) and repeated three times. (D) Measurement of apoptosis in HCEC-1CT by Annexin V staining after 24 h treatment with the compounds. Assay was performed in biological triplicates and repeated twice. p value (≤0.05), * con vs treatment; # 5-ASA vs AZTP. Kruskal-Wallis Test with Dunnet post hoc analysis was performed (E) BrdU incorporation in HCEC-1CT in the presence or absence of TNF-α (10 ng/ml) and 5-ASA (5 mM) or AZTP (10 μM). p value (≤0.05), * con vs treatment. ANOVA with Dunnett’s multiple comparisons test was performed (F) Dose dependent response of 5-ASA and AZTP (24 h) on cell cycle measured by flow cytometry using PI staining. (G) qRT-PCR analysis of genes for cell migration (CDC42) and proliferation (RR1, cyclin D1). p value (≤0.05), * con vs treatment. Dunnett’s multiple comparisons test was performed.
