Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2021 Apr 9;218(6):e20200944. doi: 10.1084/jem.20200944

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2021 Singh et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

PMC Copyright notice

Figure 7. — TNFα and TGFβ1 down-regulate DMBT1 in adjacent histologically normal epithelium. (A) Schematic for experimental design for B and C. CM from siNT or siDMBT1 HOK16B were used for the invasion assay. (B) siRNA-mediated down-regulation of DMBT1 in HOK16B. Cell lysates were immunoblotted with antibodies against ZEB1, E-cadherin (E-cad), MMP-9, and actin (n = 2). Signal intensity was quantified by densitometric analysis with normalization to actin and then expressed as percent of corresponding control (DU). (C) Scatter plots show percent invasion of UM-SCC-1 (top) and UM-SCC-29 (bottom) with CM from keratinocytes transfected with siNT or siDMBT1. Each color denotes an independent experiment with three replicates in each experiment (n = 2 for UM-SCC-1; n = 3 for UM-SCC-29). Data represent mean ± SD (*, P < 0.05; **, P < 0.01; t test). (D) Heat-map for the expression of IL10, GMCSF, TNFα, and TGFβ1 in normal (N) and cancer (T) tissues from 22 patients from GEO accession no. GSE6631. (E) Dose response of impact of recombinant TNFα and TGFβ1 on DMBT1 expression in HOK16B (n = 3). Cells were incubated with increasing concentrations of TNFα (lanes 2–5, lane 1 as vehicle control) and TGFβ1 (lanes 7–10, lane 6 as vehicle control) for 48 h. Cell lysates were immunoblotted with DMBT1 and actin antibodies. (F) Time course of recombinant TNFα and TGFβ1 treatment in HOK16B (n = 3). Cells were incubated with TNFα (10 ng/ml) or TGFβ1 (1 ng/ml). Lysates were harvested at the indicated time and immunoblotted with DMBT1 and actin antibodies. (G) HOK16B were incubated with TNFα (10 ng/ml), TGFβ1 (1 ng/ml), or vehicle. Lysates were immunoblotted with DMBT1, E-cad, MMP9, ZEB1, and actin antibodies (n = 2). (H) HOK16B transfected with siRNA (NT) or siDMBT1 were incubated with TNFα (10 ng/ml), TGFβ1 (1 ng/ml), or vehicle. Lysates were immunoblotted with DMBT1, E-cad, MMP9, ZEB1, and actin antibodies (n = 2). GFR, growth factor reduced Matrigel.