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. Author manuscript; available in PMC: 2020 Jul 11.
Published in final edited form as: Cell. 2019 Jun 27;178(2):316–329.e18. doi: 10.1016/j.cell.2019.06.003

Figure 4. Fbxo22 depletion activates Bach1 transcriptional program and promotes cell migration.

Figure 4.

(A) Volcano plot comparing the expression of Bach1 signature genes in KP cells transfected with either a non-targeting siRNA (siCtrl) or siFbxo22. Plotted for each gene are the negative log10 of the p value and the log2 of the fold change of gene expression of KP-siFbxo22 cells relative to KP-siCtrl cells. The green dots represent genes with fold change values of −/+ 2, and the red bar represents a FDR threshold of 5%. The blue dots represent down-regulated Bach1 target genes, while the red dots represent upregulated Bach1 target genes. The enrichment of transcripts was calculated considering the transcripts with at least 2 fold change at 5% FDR.

(B) The experiment was performed as in (A), except that the volcano plot compares the expression of Bach1 signature genes in KPK-siFbxo22 cells v. KPK-siCtrl cells.

(C) KP and KPK cells infected with lentiviruses expressing either a dox-inducible, non-targeting shRNA (shCtrl) or shRNAs targeting either Bach1 (shBach1) or Fbxo22 (shFbxo22). Forty-eight hours after dox induction, cells were treated with either Ki696 or TinPPIX for 24 hours. Cells were then tested for migration in a Boyden chamber assay over a 12 hours period. Next, cells migrated on the bottom of the transwells were fixed, stained and counted in 5 different fields/well. The graph shows quantification from 3 technical replicates of a representative experiment. Bottom, representative images of migrated cells. Values are presented as means ±SEM.

(D) KP-sgTom, KP-sgFbxo22, and KP-sgBach1 cells transduced with lentiviruses expressing either a dox-inducible shCtrl, shBach1, or shFbxo22 were treated with dox for 72 hours and tested for migration as in (C). Values are presented as means ±SEM.