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

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© 2021 Singh et al.

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Figure S1. — DMBT1 is down-regulated in HNSCC, and overexpression of DMBT1 suppresses invasion and tumor progression. (A) Meta-analysis showing down-regulation of DMBT1 in HNSCC relative to normal tissue. The datasets and accession numbers are given in Table S1. (B) Down-regulation of DMBT1 is correlated with poor overall survival in patients with HNSCC. The analyses were performed with the Rickman Head-Neck dataset (reporter ID, 208250_s_at; Oncomine; n = 81; *, P < 0.05; log-rank test). (C) DMBT1 gene expression in HNSCC cell lines. Total RNA from immortalized (HOK16B), primary keratinocytes (HOK5973), and HNSCC cell lines UM-SCC-(1, 14A, 29, 47, 81B, 104) was used to generate cDNAs, and Q-RT-PCR was performed. Data were analyzed by the relative quantification method with normalization to GAPDH and then relative to keratinocytes (*, P < 0.001; one-way ANOVA; error bars represent SD). Each sample was analyzed in triplicate, and the average fold-change was determined. (D) DMBT1 was stably overexpressed in UM-SCC-29 as verified by immunoblot analysis (n = 2). (E) Overexpression of DMBT1 suppresses invasion in vitro at 48 h. Each color represents an independent experiment with three replicates in each experiment (*, P < 0.05; t test). (F) Nuclear pleomorphism is significantly less in tumors with UM-SCC-29-DMBT1 cells than in control tumors with UM-SCC-29-pCMV6 cells (***, P < 0.001; t test; n = 9; error bars represent SD).