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. 2023 Jun 8;14:3375. doi: 10.1038/s41467-023-39097-x

Fig. 1. TP antibody characterization.

Fig. 1

a Representative TP-1 (left) and TP-3 (right) flow histograms for wild-type HEK-293 (top), HEK-293 transfected with ALPL-1 cDNA (HEK-293ALPL-1) (middle), and OHS cells (bottom). b Representative ALPL (left), TP-1 (middle) and TP-3 (right) flow histograms for ALPL knock out OHS (OHSALPL KO) and parental OHS. c Western Blotting of OHSALPL KO cells shown in b to confirm ALPL knock out. Source data are provided as a Source Data file. d Representative TP-1 (red) and TP-3 (blue) flow histograms for OS tumor cell lines OHS, OSA and SaOS-2. BL-41 cells (TP negative target). Secondary antibody control (grey). e Expression of ALPL-1 transcript ENST00000374840.8 in primary (n = 9) and metastatic (met) OS (n = 5), and in normal bone (n = 24) and lung tissues (n = 14). Counts were normalized by Trimmed mean of M values (TMM) method prior to log2 transformation. Statistical comparison was performed using two-tailed Mann-Whitney test. (ns, not significant p > 0.05, *p = 0.0376, **p < 0.01, ***p < 0.001). Data are presented as median with 95% confidence interval. f Flow cytometry ALPL-1 protein expression in MSCs, OS cell lines (143B, G-292, U2OS, OST-3, MG-63 and SAOS-2) and PDXs (n = 7) detected with TP-3 antibody. U2OS is open circle. Data are shown as mean ± s.d. Statistical comparisons were performed with two-tailed unpaired Student t-test (ns not significant p > 0.05, *p < 0.05, ****p < 0.0001). g Immunohistochemistry staining for ALPL-1 using TP-3 antibody on osteosarcoma (OS) and normal tissues (pancreas, stomach, uterus, lung and kidney). Images 20X, 25 micron bar.