Fig 1: AKT phosphorylation mediates TROLL-2 and TROLL-3’s functions.a Quantification of the percentage of WDR26 localization in the nuclear (Nuc) and cytoplasmic (Cyt) fractions of CA1D cells transfected with the indicated siRNAs. Data are mean ± SD and analysed with two-way ANOVA. n = 3 biological replicates, asterisk vs. siControl, P < 0.05. b Representative western blot analysis of the coimmuno-precipitation of endogenous NOLC1 and WDR26 in MCF10A cells transfected with the indicated constructs and siRNAs. c Representative western blot of WDR26 and NOLC1 localization in the nuclear (Nuc) and cytoplasmic (Cyt) fractions of MCF10A cells transfected with the indicated siRNAs. d Quantification of cell migration of CA1D cells overexpressing either TROLL-2, TROLL-3, or empty vector, in combination with the indicated WDR26 constructs and transfected with the indicated siRNAs. Data are mean ± SD and analysed with two-tailed Student’s t test, n = 3 biological replicates, asterisk vs. pBabe Empty siControl, P < 0.005. Section vs. pBabe TROLL-2 siControl, P < 0.005. Hash vs. pBabe TROLL-3 siControl, P < 0.005. e Representative western blot analysis of the coimmuno-precipitation of endogenous AKT and WDR26 in CA1D cells transfected with the indicated siRNAs. f–h Representative H&E images (f) and IHC for WDR26 (g) and for pAKT (h) in tumours derived from DCIS cells infected with the indicated constructs (n = 5 mice for both groups). All blots are representative of n = 3 biological replicates. Source data are provided as Source data file.
Fig 2: Model. Scheme describing the proposed mechanism of action of TROLL-2 and TROLL-3.In normal cells (e.g. MCF10A cells) the tumour and metastasis suppressor TAp63 inhibits the expression of TROLL-2 and TROLL-3, while NOLC1 interacts with WDR26 and promotes the accumulation of WDR26 in the nucleus. In cancer cells (e.g. CA1D cells), instead, mutant p53 inhibits TAp63, thus allowing for the expression of TROLL-2 and TROLL-3. These lncRNAs counteract the interaction between NOLC1 and WDR26, while promoting the binding of WDR26 to AKT. As a consequence, the PI3K/AKT pathway is activated and can sustain tumour formation and progression.
Fig 3: TROLL-2 and TROLL-3 promote cancer progression via WDR26.a Venn diagram of the proteins interacting with TROLL-2 and TROLL-3. b Table listing the 7 common interactors of TROLL-2 and TROLL-3. c, d Quantification of cell migration (c) and invasion (d) of CA1D cells overexpressing either TROLL-2, TROLL-3, or the empty vector, and transfected with the indicated siRNAs. Graphs represent the individual data points, mean ± SD and analysed with two-tailed Student’s t test, n = 3 biological replicates, asterisk vs. pBabe Empty siControl, P < 0.005. Section vs. pBabe TROLL-2 siControl, P < 0.005. Hash vs. pBabe TROLL-3 siControl, P < 0.005. e Representative western blot for the indicated proteins pulled down by the indicated lncRNAs. n = 3 biological replicates. Source data are provided as Source data file.
Fig 4: Cytoplasmic WDR26 correlates with advanced cancers.a Circos plot summarizing the expression of TROLL-2, TROLL-3, WDR26 and pAKT in TMAs of 378 cancers with progressive disease. b Representative H&E stained cross sections of melanomas derived from Malme-3M cells infected with the indicated shRNAs. c Quantification of the melanomas described in b. n = 5 mice for all groups, asterisk vs. NT, P < 0.005, two-tailed Student’s t test. d Representative H&E stained cross sections of lung colonies derived from Malme-3M cells infected with the indicated shRNAs. e Quantification of the lung colonies described in d. n = 5 mice for all groups, asterisk vs. NT, P < 0.005, two-tailed Student’s t test. Boxplots represent the individual data points, median and whiskers (min to max method). Source data are provided as Source data file.
Fig 5: WDR26 localization correlates with breast cancer progression.a Representative IHC images for WDR26 in lobular hyperplasia (left) and invasive ductal carcinoma (right). b Quantification of the percentage of WDR26 cellular distribution in the TMA of breast cancer progression shown in Fig. 2a. c–f Correlation of the IHC score and cellular distribution of WDR26 with the ISH score of TROLL-2 and TROLL-3 in the indicated TMAs based on tissue type (c, d) or tumour grade (e, f). Source data are provided as Source data file.
Supplier Page from Abcam for Anti-WDR26 antibody