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. 2020 Dec 24;4(2):e202000805. doi: 10.26508/lsa.202000805

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© 2020 Dunbar et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure 5. — (A) qRT-PCR was performed using cDNA from HCT116 wild-type and HCT116 FAM83F^−/− (cl.1) cell lines following treatment with L-CM or Wnt3A-CM with or without 0.5 μM CHIR99021 for 6 h, and primers for Axin2 and GAPDH genes. Axin2 mRNA expression was normalised to GAPDH mRNA expression and represented as arbitrary units. Data presented as scatter graph illustrating individual data points with an overlay of the mean ± SD. (B) Cytoplasmic, nuclear, and membrane lysates from HCT116 wild-type, HCT116 FAM83F^−/− (cl.1), and HCT116 FAM83F^−/− (cl.2) cell lines were resolved by SDS–PAGE and subjected to Western blotting with indicated antibodies. (C) Densitometry of CK1α protein abundance from (B) membrane lysates normalised to GAPDH protein abundance and represented as fold change compared with HCT116 wild-type cells. Data presented as scatter graph illustrating individual data points with an overlay of the mean ± SD. (D) Cytoplasmic, nuclear, and membrane lysates from DLD-1 wild-type and DLD-1 FAM83F^−/− cell lines were resolved by SDS–PAGE and subjected to Western blotting with indicated antibodies. (E) Densitometry of CK1α protein abundance from (D) membrane lysates normalised to Na/K ATPase protein abundance and represented as fold change compared with DLD-1 wild-type cells. Data presented as scatter graph illustrating individual data points with an overlay of the mean ± SD. (B, D) The specificity of cytoplasmic, nuclear and membrane compartment lysates were determined by Western blotting with the following subcellular compartment-specific antibodies: α-tubulin (cytoplasmic), Lamin A/C (nuclear), U2AF1 (nuclear), and Na/K ATPase (membrane). (A, C, E) Statistical analysis was completed using a Student’s unpaired t test and comparing cell lines as denoted on graphs. Statistically significant P-values are denoted by asterisks (**** < 0.0001, *** < 0.001, ** < 0.01, * < 0.05).

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