Fig 1: The effects of PIP on OSCC migration and invasion in vitro. (a) The view of wound-healing migration assay; the average rate of SCC15 and SCC25 cells migration at 12 and 24 hr after control, PIP vector or siRNA-PIP transfection. All data were shown as mean ± SD. *p < 0.05, **p < 0.01. (b) The results of the transwell assay of SCC15 and SCC25 cells at 24 hr after control, PIP vector or siRNA-PIP transfection; relative ratios of migrated and invasive cells per field are shown. All data were shown as mean ± SD. *p < 0.05, **p < 0.01. OSCC: oral squamous cell carcinoma; PIP: prolactin-inducible protein; SD: standard deviation; siRNA: small interfering RNA [Color figure can be viewed at wileyonlinelibrary.com]
Fig 2: Effects of siRNA-PIP coculture and AKT/MAPK inhibitor on OSCC cell proliferation. (a) Cells treated with DMSO, MK-2206, SCH772984, siRNA-PIP + MK-2206 or siRNA-PIP + SCH772984 were assessed by colony forming assays as described in Figure 2(a). All data were shown as mean ± SD. *p < 0.05, **p < 0.01. (b) At 24, 48, and 72 hr posttreatment with DMSO, MK-2206, SCH772984, siRNA-PIP + MK-2206 or siRNA-PIP + SCH772984, cell proliferation was examined via MTT assays. (c) Cell cycle analysis of SCC15 and SCC25 cell cycle treated as in (a). All data were shown as mean ± SD. *p < 0.05, **p < 0.01. AKT/MAPK: Akt/mitogen-activated protein kinase; DMSO: dimethyl sulfoxide; MTT: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide; OSCC: oral squamous cell carcinoma; PIP: prolactin-inducible protein; SD: standard deviation; siRNA: small interfering RNA [Color figure can be viewed at wileyonlinelibrary.com]
Fig 3: PIP detection in clinical specimens. (a) Molecular function of Gene Ontology (GO) analyses. (b) List of DEPs with binding by iTRAQ analysis; ten DEPs (ACTN1, APOBEC3G, CASQ1, DPYSL2, FLG, FLG2, PIP, SDCBP2, SDF4, TPM1) were assessed via qRT-PCR analysis in OSCC and adjacent normal tissues (n = 29). (c) qRT-PCR analysis of PIP expression in OSCC and adjacent normal tissues (n = 29); western blot analysis of PIP expression in OSCC and adjacent normal tissues (n = 5). The expression of PIP was normalized to ß-actin. Data were from three independent experiments and represented as mean ± SD. *p < 0.05. DEP: differentially expressed protein; iTRAQ: isobaric tags for relative and absolute quantitation; OSCC: oral squamous cell carcinoma; PIP: prolactin-inducible protein; qRT-PCR: quantitative real-time polymerase chain reaction; SD: standard deviation [Color figure can be viewed at wileyonlinelibrary.com]
Fig 4: PIP expression in OSCC tissues. Immunohistochemistry (IHC) analysis of PIP in OSCC and adjacent normal tissues (n = 45; ×20). OSCC: oral squamous cell carcinoma; PIP: prolactin-inducible protein [Color figure can be viewed at wileyonlinelibrary.com]
Fig 5: PPI network suggests that PIP interacts with numerous components of the AKT/MAPK pathway. (a) PPI network for PIP was retrieved from the STRING database and reconstructed by adding recently identified PIP interacting proteins. (b) Western blot analyses in SCC15 and SCC25 cells after transfection with control, PIP vector, and siRNA-PIP. ß-Actin served as an internal control. (c) Proposed model for the effects of PIP on OSCC cell proliferation, invasion, migration and AKT/MAPK signaling. AKT/MAPK: Akt/mitogen-activated protein kinase; OSCC: oral squamous cell carcinoma; PIP: prolactin-inducible protein; PPI: protein–protein interaction; siRNA: small interfering RNA [Color figure can be viewed at wileyonlinelibrary.com]
Supplier Page from Abcam for Anti-GCDFP 15 antibody [PIP/1571]