Figure 2
Comparative analysis of hCPC nuclear compartment. (a) Ingenuity pathway analysis (IPA) of the nuclear hCPC subproteome obtained by whole label-free (LF) proteomics; (b), (c). Main overexpressed (b) or downregulated (c) proteins in purified nuclear fractions of hCPC compared with hMSC, analyzed by LF proteomics (n = 3); code: + + + , indicates 5–10 peptides; + + , 2–4 peptides; + , 1 peptide, + / − , 0–1 peptide and –-, no peptide detected. In parallel, comparative levels of the indicated proteins in cytoplasmic extracts are also indicated. (d) PANTHER GO-Slim Biological Processes analysis of overexpressed nuclear proteins in hCPC. (e) Comparative RT-qPCR expression analysis of ASPHD1 in the three independent isolates of hCPC (hCPC 1–3), two human fibroblasts (HDF1 and HDF2) and two hMSC isolates (MSC19 and MSC45). Assays were performed three times and data are expressed as mean ± SD; black lines summarize p-values (*** < 0.002) for hCPC vs. fibroblasts or hMSC (One-way ANOVA analysis of variance followed by the Bonferroni correction for multiple comparison). (f) Western blot analysis of ASPHD1 in the three independent isolates of hCPC (hCPC 1–3), human dermal fibroblasts (HDF1) and hMSC (MSC19) isolate; tubulin was used as a loading control.'Full-length blots/gels' are presented in Supplementary Figure S4 online. (g) Comparative immunofluorescence analysis of ASPHD1 (red) in hCPC (hCPC 1 and 3), hMSC (MSC19) and human dermal fibroblasts (HDF); nuclei were counterstained with DAPI (blue). Bar, 20 μm.
