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. Author manuscript; available in PMC: 2021 Oct 15.
Published in final edited form as: Cancer Res. 2021 Feb 8;81(8):2171–2183. doi: 10.1158/0008-5472.CAN-20-1458

Figure 2: USP6-regulated chemokines are induced by Jak-STAT1/NF-κB and can stimulate immune cell migration.

Figure 2:

A) USP6/RD-ES with the indicated CRISPR deletions were treated with dox and IFNγ (50 ng/ml), and CXCL9/10 expression was quantified. B) USP6(Pool)/RD-ES were treated with dox and IFNγ overnight, in the presence of PS1145 (15 μM) and Jak inhibitor (1 μM) as indicated, and CXCL9/10 expression quantified. C) Firefly (FF) luciferase driven by the CXCL10 promoter (WT or mutants in the indicated response elements) were co-transfected with renilla luciferase and USP6-encoding vectors. Cells were treated overnight with IFNγ, then subjected to dual luciferase assays. FF/renilla ratios relative to WT (black bar) were calculated; all p values are in comparison to this sample. Transwell migration assays were performed on THP-1 (D), and primary monocytes (E). Migration was monitored after 2 hr in response to serum-free conditioned medium (CM) from USP6/RD-ES cells treated as indicated, control serum-free medium, or recombinant CCL5 (10 ng/ml), CXCL10 (250 ng/ml), or 1% fetal bovine serum (FBS).