Journal: Cellular signalling

Article Title: Casein kinase 1 controls the shuttling of epidermal growth factor receptor and estrogen receptor in endometrial carcinoma induced by breast cancer hormonal therapy: Relevance of GPER1/Src.

doi: 10.1016/j.cellsig.2023.110733

Figure Lengend Snippet: Fig. 2. Inhibitory effects of CKI-7/IC261 on the translocation and expression of EGFR and ERα in 4-OHT-treated Ishikawa cells. (A-B) Representative immunoblots for showing changes in p-EGFR, EGFR, p-ERαS118, p-ERαS167, and ERα levels in Ishikawa cells treated with 1 μM 4-OHT with or without (A) 1 μM CKI-7 and (B) 0.05 nM IC261 for 48 h. α-Tubulin was used as the control. The protein expression levels, as determined by individual bands of the phosphorylated form, were compared to the expression levels of α-tubulin and/or the total protein (pan protein) (n = 4). (C) Immunofluorescence images of nuclear ERα expression in Ishikawa cells treated with 4-OHT with or without 1 μM CKI-7 or 0.05 nM IC261 for 48 h. Cells were labeled using an anti-ERα antibody (red) and the nuclear stain DAPI (blue). Images were captured using a confocal microscope (LSM800, Zeiss). Scale bar: 20 μm. (D) Immunofluorescence images of EGFR translocation and expression at 0 h, 3 h, and 48 h in Ishikawa cells treated with 4-OHT with or without 1 μM CKI-7 or 0.05 nM IC261. Cells were labeled using an anti-EGFR antibody (red) and the nuclear stain DAPI (blue). Scale bar: 20 μm. (E) Representative immunoblots for EGFR localization at 0 h, 3 h, and 48 h in subcellular fractions of Ishikawa cells treated with 1 μM 4-OHT with or without 1 μM CKI-7 or 0.05 nM IC261. Na+/K+ATPase, GAPDH, and histone H3 were used as markers of the plasma membrane, cytoplasm, and nucleus, respectively. The protein expression levels, as determined by individual bands, were respectively compared to the expression levels of Na+/K+ATPase, GAPDH, and histone H3 (n = 4). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Antibodies against HER2 (#2242), progesterone receptor (PR; #8757), phospho-ERα (p-ERα) (ERαSer167, #64508; p-ERαSer118, #2511), EGFR (D38B1) XP® (#4267), p-EGFRTyr1068 (D7A5) XP® (#3777), Src (#2109), p-SrcTyr416 (#6943), Histone H3 (D1H2) (#4499), GAPDH (D16H11) XP® (#5174), and Na,K-ATPase α1 (D4Y7E) (#23565) were purchased from Cell Signaling (Danvers, MA, USA).

Techniques: Translocation Assay, Expressing, Western Blot, Control, Immunofluorescence, Labeling, Staining, Microscopy, Clinical Proteomics, Membrane

Journal: Cellular signalling

Article Title: Casein kinase 1 controls the shuttling of epidermal growth factor receptor and estrogen receptor in endometrial carcinoma induced by breast cancer hormonal therapy: Relevance of GPER1/Src.

doi: 10.1016/j.cellsig.2023.110733

Figure Lengend Snippet: Fig. 5. Validation of signaling pathways associated with 4-OHT-induced EGFR and ERα translocation and expression in endometrial cells. (A-B) Immunofluorescence images of ERα (A) and EGFR (B) localization and expression in Ishikawa cells subjected to different treatments for 48 h: vehicle, 1 μM 4-OHT with or without 1 μM Gef, 0.1 μM G15, and 0.1 μM PP1. Cells were labeled using an anti- ERα antibody (red), EGFR (green), and the nuclear stain DAPI (blue). Images were captured using a confocal microscope (LSM800, Zeiss). Scale bar: 10 μm. (C) Representative immunoblots of EGFR localization at 0 h, 3 h, and 48 h in subcellular fractions of Ishikawa cells treated with 1 μM 4-OHT with or without 1 μM Gef, 0.1 μM G15, or 0.1 μM PP1. Na+/K+ATPase, GAPDH, and histone H3 were used as markers of the plasma membrane, cytoplasm, and nucleus, respectively. The protein expression levels, as determined by individual bands, were respectively compared to the expression levels of Na+/K+ATPase, GAPDH, and histone H3 (n = 4). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Antibodies against HER2 (#2242), progesterone receptor (PR; #8757), phospho-ERα (p-ERα) (ERαSer167, #64508; p-ERαSer118, #2511), EGFR (D38B1) XP® (#4267), p-EGFRTyr1068 (D7A5) XP® (#3777), Src (#2109), p-SrcTyr416 (#6943), Histone H3 (D1H2) (#4499), GAPDH (D16H11) XP® (#5174), and Na,K-ATPase α1 (D4Y7E) (#23565) were purchased from Cell Signaling (Danvers, MA, USA).

Techniques: Biomarker Discovery, Protein-Protein interactions, Translocation Assay, Expressing, Immunofluorescence, Labeling, Staining, Microscopy, Western Blot, Clinical Proteomics, Membrane