Fig. 1: G007-LK can reduce WNT/β-catenin signaling in B16-F10 cells in vitro. | Communications Biology

Fig. 1: G007-LK can reduce WNT/β-catenin signaling in B16-F10 cells in vitro.

From: Tankyrase inhibition sensitizes melanoma to PD-1 immune checkpoint blockade in syngeneic mouse models

Fig. 1

a Representative immunoblots of cytoplasmic AXIN1 (upper) and nuclear active form of β-catenin (non-phospho, serine [Ser] 33/37/threonine [Thr] 41) and total β-catenin (lower). GAPDH or lamin B1 document equal protein loading. Treatments used for cultured B16-F10 cells in a–c: Vehicle (DMSO, 0.01%), G007-LK (1 µM), recombinant WNT3a (activator of WNT/β-catenin signaling) or WNT3a + G007-LK for 24 h. b Luciferase-based reporter assay for measuring WNT/β-catenin signaling activity. B16-F10 cells transiently transfected with superTOPflash (vector with TCF promoter binding sites) or FOPflash (control vector with mutated TCF binding sites) along with Renilla luciferase (for normalization). All samples normalized to superTOPflash signal for wild-type control. For b, c Boxplots show median, first and third quartiles and maximum and minimum whiskers. One-tailed t-tests are indicated by *(P < 0.05) or **(P < 0.01). Background SuperTOPflash versus FOPflash activities were not significantly different, indicating low basal WNT/β-catenin signaling activation. One representative experiment of two repeated assays with three replicates is shown. c Real-time RT-qPCR analyses of WNT/β-catenin signaling target genes (Axin2 and Tcf7). One-tailed t-test is indicated by **(P < 0.01) and one-tailed Mann-Whitney rank sum tests are indicated by †(P < 0.05). Combined data from minimum three independent experiments with three replicates each are shown.

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