vtRNA1-1 induced mitoxantrone independent with the existence of MVP.

<p>(A) The efficiency of MVP or vtRNA1-1 knockdown was detected by semiquantitative western blot or qRT-PCR. (B) DOX sensitivity to MCF-7 cells with or without vtRNA1-1 knockdown was detected. (C) Immunofluorescence imaging was performed to detect the sub-cellular localization of DOX with or without vtRNA1-1 knockdown.</p

vtRNA1-1 promotes the tumor formation and enhances the chemoresistance of DOX in vitro.

<p>(A) After transfection of vtRNA1-1, RBD or vtRNA1-1/RBD, sensitivity to DOX was measured in MCF-7 cells. (B) the expressing levels of GAGE6 and PSF after the DOX treatment were detected in MCF-7 transfected with vtRNA1-1, RBD or vtRNA1-1/RBD and the relative folds were qualified (C). (D) ChIP was performed to detect the binding of PSF to GAGE6 promoter in MCF-7 transfected with vtRNA1-1, RBD or vtRNA1-1/RBD after DOX exposure. CCK-8 assay (E) or PI staining (F) was performed to detect cell viability in MCF-7 transfected with vtRNA1-1 after DOX exposure. *P<0.05.</p

VRNA binds to PSF on its RNA binding domain (RBD) and releases GAGE6 promoter from PSF in vitro.

<p>(A) Northern blot was performed to detect the existence of vtRNA1-1, 1–2, 1–3 or 2–1. (B) qRT-PCR analysis was performed to quantify the relative amount of vtRNA1-1, 1–2, 1–3 or 2–1 normalized to 5.8S rRNA. (C) EMSA analysis was performed the binding of vtRNA1-1, 1–2, 1–3 or 2–1 to purified PSF in vitro. (D) The specific binding of vtRNA1-1 to RBD of PSF was detected. (E) the competitive binding of vtRNA1-1 to PSF against GAGE6 promoter fragment was analyzed.</p

The effects of vtRNA1-1 in cell proliferation and colony formation in a PSF-dependent way.

<p>Cell curve assays were performed to identify the association of vRNA with PSF (A, left panel), RBD or DBD (B, right panel) on affecting the cell proliferation in MCF-7. (B) EdU staining was performed to further confirm the effect of vRNA on cell proliferation. (C) Soft agar assay was performed to present vRNA’s effect on colony formation in MCF-7. *P<0.05.</p

vtRNA1-1 expression tightly regulates the expressing level of GAGE6 in MCF-7 and MCF-7/MR.

<p>(A) RNA-IP assay confirmed the specific binding of PSF to vtRNA1-1 through it RBD in vivo. RT-qPCR (B) and semi-quantitative western blot (C) presents the expressing changes in the mRNA and protein level of GAGE6 in MCF-7 and MCF-7/MR. Chromatin immunoprecipitation assays present that the binding capacity of PSF to GAGE6’s promoter region affected by vtRNA1-1 in MCF-7 (D) or MCF-7/MR (E). RT-qPCR (F) and Semiquantitative western blot (G) present the mRNA and protein levels affected by the ectopic expression of vtRNA1-1 in MCF-7 or MCF-7/MR. *P<0.05.</p

Quantification of the staining intensity for E-cadherin, ZO-1 and occludin in control and mature CRSwNP.

<p>Quantification of the staining intensity for E-cadherin, ZO-1 and occludin in control and mature CRSwNP.</p

Epithelial loss (%) in control, mature CRSwNP and middle turbinate CRSwNP.

<p>MT, middle turbinate.</p

Quantification of celluar and extracelluar components.

<p>Eosinophils(A), CD68-(B), MMR-(C), α-SMA-(D), vimentin-(E), and pSmad2(F) positive cells, Fn (G) and collagen(H) in control interior turbinates (control IT), mature CRSwNP and middle turbinate CRSwNP, separated into turbinate and polyp areas. </p

Immunostaining of celluar and extracelluar components.

<p>HE (A1-A5), CD68 (B1-B5),MMR (C1-C5), α-SMA (D1-D5), vimentin (E1-E5), pSmad2 (F1-F5), Fn (G1-G5) picrosirius red staining viewed in bright-field (H1-H5) and viewed under polarized light (I1-I5) in mature CRSwNP (A1-G1), polyp area of middle turbinate CRSwNP (A2-G2),turbinate area of middle turbinate CRSwNP (A4-G4) and control mucosa (A5-G5)(final magnification 400×). The overview of middle turbinate CRSwNP was shown in A3-G3 (composed by Photoshop software with pictures in 40×magnification).</p

Fn mRNA expression and Fn protein concentration.

<p>Fn mRNA expression and Fn protein concentration.</p