Fig 2: Anti-tumor effect of XPC on A549 tumor-bearing nude mice. (A). Xenografts were generated by A549 cells (shCtrl), A549 cells with stable XPC knockdown (shXPC). Tumors were removed from mice after 32 days. (B). Tumor sizes were measured every 4 days. (C). The expression of CD133 and OCT4 were determined using Flow Cytometry. p < 0.0001(CD133), p < 0.0001(OCT4), A549-shXPC compared with A549-shCtrl. (D). The expression of XPC and CD133 were determined using Western blotting. The intensity of each band was quantified using Image J and normalized to. Protein extractions come from xenotransplanted tumor tissue of mice. ***p < 0.001 (XPC), *p < 0.05(CD133), A549-shXPC compared with A549-shCtrl. (n = 6).

Fig 3: Expression of trilineage differentiation markers. A) Matched iPSCs and differentiated endoderm, mesoderm, ectoderm cells were analyzed by immunostaining followed by flow cytometry for expression of markers specific for each germ layer (mean ± SEM, n = 7–10 cell lines). See also Fig. S1 for isotype control data, Fig. S3 for morphological changes after differentiation, Fig. S4 for paired data, and Fig. S5 for independent validation of iPSC NES staining. B) Representative flow cytometry plots showing co-immunostaining of pluripotency marker POU5F1 with each of the four candidate ectoderm markers in undifferentiated iPSCs. See also Fig. S6 for complete dataset. C, D) Expression of pluripotent marker POU5F1 (C) and four candidate ectoderm markers (D) quantified by qPCR in matched iPSCs and ectoderm differentiated cells (mean ± SEM, n = 8). Gene expression is shown as fold change of ectoderm differentiated cells compared to iPSCs, with dotted line indicating fold change of 1. * P < .05, *** P < .001, **** P < .0001, two-tailed paired t-tests.

Fig 4: Validation of blood-derived human iPSCs. Live stain of representative iPSC colonies showing A) alkaline phosphatase activity, and B) SSEA-4 expression. C) Flow cytometry analysis of pluripotency markers POU5F1 and TRA-1–60, and differentiation marker SSEA-1 in iPSCs (mean ± SD, n = 30). See also Fig. S1 for flow cytometry data on isotype controls. D) Karyotype analysis of three iPSC lines showed normal chromosomal content. See also Fig. S2 for additional characterization of pluripotency marker expression and genomic aberrations.

Fig 5: The effect of XPC expression on stem cell markers. (A). The 3 cell lines A549, H1299, and H1650 were divided into two groups: siCtrl group and siXPC group. Western blot was used to detect the expression of stem cell markers in the siCtrl group and siXPC group. (B). In the 3 cell lines A549, H1299, and H1650, the expression of XPC in the siXPC group was lower than that in the siCtrl group (**p < 0.01, ***p < 0.001, ****p < 0.0001). (C). In the A549 and H1650 cell lines, the expression of OCT4 in the siXPC group was higher than that in the siCtrol group (*p < 0.05, ****p < 0.0001). (D). In the 3 cell lines A549, H1299, and H1650, the expression of CD133 in the siXPC group was higher than that in the siCtrl group (*p < 0.05, **p < 0.01). (E,F). The protein levels of CD133 and OCT4 were determined using Flow Cytometry. p < 0.0001(CD133), p < 0.0001(OCT4), A549-shXPC compared with A549-shCtrl. (G). Immunohistochemical detection of XPC and CD133 expression correlation in 140 cases of lung adenocarcinoma tissues (p < 0.0006, R = −0.2580).