Fig. 1
From: Differential expression of Cosmc, T-synthase and mucins in Tn-positive colorectal cancers
Loss of function of Cosmc in LS 180-Tn(+) and HCT8-Tn(+) cells. a, immunofluorescence of the Tn antigen in LS 180 parental, Tn(ā) and Tn(+) cells. LS 180 parental cells contained a small number of Tn-positive cells (green). Compared to LS 180-Tn(ā) cells, Tn(+) cells expressed robust cell surface Tn antigen (red). b, immunofluorescence of the Tn antigen in HCT8 parental, Tn(ā) and Tn(+) cells. Compared to HCT8 parental and Tn(ā) cells, Tn(+) cells expressed cell surface Tn antigen (green). In a and b, nuclei were counterstained with DAPI (blue), all scale bars are 50Ā Ī¼m. c, flow cytometry analyses of LS 180 and HCT8 subpopulations. Histograms of parental, Tn(ā) and Tn(+) cells are shown as red, blue and green lines, respectively. Inset numbers show %Tn(+), defined by the horizontal grey gate. d, sequencing analyses of Cosmc in LS 180 and HCT8 cells. Parental and Tn(ā) cells contained WT Cosmc coding region, while LS 180-Tn(+) and HCT8-Tn(+) cells had single nucleotide deletion at nt473 and nt482, respectively. Cell line names are on top, nucleotide positions are labeled above the trace files. The delT473 and delA482 are indicated by arrows. e, enzyme activities of T-synthase in LS 180 and HCT8 subpopulations. Compared to the parental and Tn(ā) cells, Tn(+) cells had significantly lower enzyme activity of T-synthase. Activity values were determined in triplicates, error bars represent the standard error of the mean (SEM). f, expression of the Tn antigen, T-synthase, Cosmc, and Ī±-tubulin in LS 180 and HCT8 cells. LS 180-Tn(+) cells expressed significant amounts of the Tn antigen, whereas HCT8-Tn(+) cells exhibited slightly increased Tn antigen (arrows). There was no detectable T-synthase and Cosmc proteins in both LS 180-Tn(+) and HCT8-Tn(+) cells. Although Ī±-tubulin levels varied in HCT8 subpopulations, Ponceau S staining indicated equal amount of total proteins loaded for WB. Names of cell populations are listed on top. Protein standards are labeled at left, and antibodies at right