Journal: JCI Insight

Article Title: Epithelial HO-1 regulates iron availability and promotes colonic tumorigenesis in a context-dependent manner

doi: 10.1172/jci.insight.181032

Figure Lengend Snippet: ( A ) Representative photomicrographs of immunofluorescence staining for 8-hydroxy-2′-deoxyguanosine (8-OHdG) and Ki-67, using paraffin-embedded mouse colonic individual tumor sections from 4 control and 4 Hmox1 ΔIEC mice undergoing AOM-DSS (original magnification, ×10). ( B ) Quantification of 8-OHdG staining intensity in tumor tissue. ( C ) 8-OHdG staining intensity in adjacent nontumor colonic tissue from control and KO mice. ( D ) Percentage of Ki-67 + cells based on immunofluorescence staining. ( E ) Ki-67 H-score analysis from tumor sections. ( F ) Representative photomicrographs of immunofluorescence staining for 4-HNE and EpCAM using colon tumor sections (original magnification, ×10). ( G ) Quantification of 4-HNE from tumor sections taken from control and KO mouse tumors. ( H ) Whole tumor iron concentration measured by colorimetric assay. ( I ) 4-HNE staining intensity normalized to mean tumor iron levels per genotype. ( J ) Tumoroids derived from pooled tumors from 2 Hmox1 fl/fl mice (control A and B ) and 1 Hmox1 ΔIEC (KO) mice were exposed to hemin (100 μM) for 24 hours. Lipid peroxidation measured by flow cytometry for 4-HNE, with median fluorescence intensity (MFI) values normalized to the mean of each vehicle-treated group. Data are shown as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, and *** *P < 0.0001 by unpaired, Student’s t tests ( B - E, G - I ) and ANOVA ( J ) with correction for multiple comparisons.

Article Snippet: The Iron Assay Kit (Colorimetric) from Novus Biologicals was used to measure total iron content.

Techniques: Immunofluorescence, Staining, Control, Concentration Assay, Colorimetric Assay, Derivative Assay, Flow Cytometry, Fluorescence