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Fig. 6 Ezrin and NHERF1 are required for <t>Erbin</t> and HER2 interactions. A) Immunofluorescence for Erbin with HER2 (top row), NHERF1 (middle row), and Ezrin (bottom row) in NSC668394 treated SKBR3 cells. All scale bars represent 10µM. B) Immunofluorescence for Erbin with HER2 (top row) and Ezrin (bot tom row) in NHERF1 knockdown SKBR3 cells. All scale bars represent 10µM. C) PLA fluorescence showing protein-protein interactions between HER2/ Erbin, Erbin/NHERF1, or Erbin/Ezrin in control, NSC668394 treated-, and NHERF1 knock-down SKBR3 cells. All scale bars represent 10µM. (n = 3) D) Quan tification of PLA results was performed by measuring the fluorescent intensity of PLA signals at the cell surface. HER2/Erbin (control n = 14, NSC668394 n = 14, NHERF1KD n = 14), Erbin/NHERF1 (control n = 19, NSC668394 n = 19, NHERF1KD n = 19), Erbin/EZRIN (control n = 15, NSC668394 n = 15, NHERF1KD n = 15). Bar graphs represent the mean ± SEM. **** denotes p < 0.00005. (n = 3). E) A working model proposing how HER2 mediated disruption of apical/ basal polarity creates the opportunity for the formation of a multi-protein signaling complex that includes HER2, Erbin, PMCA2, NHERF1, and Ezrin. Cre ated with BioRender.com
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Fig. 6 Ezrin and NHERF1 are required for <t>Erbin</t> and HER2 interactions. A) Immunofluorescence for Erbin with HER2 (top row), NHERF1 (middle row), and Ezrin (bottom row) in NSC668394 treated SKBR3 cells. All scale bars represent 10µM. B) Immunofluorescence for Erbin with HER2 (top row) and Ezrin (bot tom row) in NHERF1 knockdown SKBR3 cells. All scale bars represent 10µM. C) PLA fluorescence showing protein-protein interactions between HER2/ Erbin, Erbin/NHERF1, or Erbin/Ezrin in control, NSC668394 treated-, and NHERF1 knock-down SKBR3 cells. All scale bars represent 10µM. (n = 3) D) Quan tification of PLA results was performed by measuring the fluorescent intensity of PLA signals at the cell surface. HER2/Erbin (control n = 14, NSC668394 n = 14, NHERF1KD n = 14), Erbin/NHERF1 (control n = 19, NSC668394 n = 19, NHERF1KD n = 19), Erbin/EZRIN (control n = 15, NSC668394 n = 15, NHERF1KD n = 15). Bar graphs represent the mean ± SEM. **** denotes p < 0.00005. (n = 3). E) A working model proposing how HER2 mediated disruption of apical/ basal polarity creates the opportunity for the formation of a multi-protein signaling complex that includes HER2, Erbin, PMCA2, NHERF1, and Ezrin. Cre ated with BioRender.com
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lentiviruses containing three individual shrna targeting erbin mrna sc-40541-v  (Santa Cruz Biotechnology)
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Fig. 6 Ezrin and NHERF1 are required for <t>Erbin</t> and HER2 interactions. A) Immunofluorescence for Erbin with HER2 (top row), NHERF1 (middle row), and Ezrin (bottom row) in NSC668394 treated SKBR3 cells. All scale bars represent 10µM. B) Immunofluorescence for Erbin with HER2 (top row) and Ezrin (bot tom row) in NHERF1 knockdown SKBR3 cells. All scale bars represent 10µM. C) PLA fluorescence showing protein-protein interactions between HER2/ Erbin, Erbin/NHERF1, or Erbin/Ezrin in control, NSC668394 treated-, and NHERF1 knock-down SKBR3 cells. All scale bars represent 10µM. (n = 3) D) Quan tification of PLA results was performed by measuring the fluorescent intensity of PLA signals at the cell surface. HER2/Erbin (control n = 14, NSC668394 n = 14, NHERF1KD n = 14), Erbin/NHERF1 (control n = 19, NSC668394 n = 19, NHERF1KD n = 19), Erbin/EZRIN (control n = 15, NSC668394 n = 15, NHERF1KD n = 15). Bar graphs represent the mean ± SEM. **** denotes p < 0.00005. (n = 3). E) A working model proposing how HER2 mediated disruption of apical/ basal polarity creates the opportunity for the formation of a multi-protein signaling complex that includes HER2, Erbin, PMCA2, NHERF1, and Ezrin. Cre ated with BioRender.com
Lentiviruses Containing Three Individual Shrna Targeting Erbin Mrna Sc 40541 V, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Fig. 6 Ezrin and NHERF1 are required for Erbin and HER2 interactions. A) Immunofluorescence for Erbin with HER2 (top row), NHERF1 (middle row), and Ezrin (bottom row) in NSC668394 treated SKBR3 cells. All scale bars represent 10µM. B) Immunofluorescence for Erbin with HER2 (top row) and Ezrin (bot tom row) in NHERF1 knockdown SKBR3 cells. All scale bars represent 10µM. C) PLA fluorescence showing protein-protein interactions between HER2/ Erbin, Erbin/NHERF1, or Erbin/Ezrin in control, NSC668394 treated-, and NHERF1 knock-down SKBR3 cells. All scale bars represent 10µM. (n = 3) D) Quan tification of PLA results was performed by measuring the fluorescent intensity of PLA signals at the cell surface. HER2/Erbin (control n = 14, NSC668394 n = 14, NHERF1KD n = 14), Erbin/NHERF1 (control n = 19, NSC668394 n = 19, NHERF1KD n = 19), Erbin/EZRIN (control n = 15, NSC668394 n = 15, NHERF1KD n = 15). Bar graphs represent the mean ± SEM. **** denotes p < 0.00005. (n = 3). E) A working model proposing how HER2 mediated disruption of apical/ basal polarity creates the opportunity for the formation of a multi-protein signaling complex that includes HER2, Erbin, PMCA2, NHERF1, and Ezrin. Cre ated with BioRender.com

Journal: Breast cancer research : BCR

Article Title: Erbin interacts with NHERF1 and Ezrin to stabilize a membrane ErbB2 signaling complex in HER2-positive breast cancer.

doi: 10.1186/s13058-025-02025-6

Figure Lengend Snippet: Fig. 6 Ezrin and NHERF1 are required for Erbin and HER2 interactions. A) Immunofluorescence for Erbin with HER2 (top row), NHERF1 (middle row), and Ezrin (bottom row) in NSC668394 treated SKBR3 cells. All scale bars represent 10µM. B) Immunofluorescence for Erbin with HER2 (top row) and Ezrin (bot tom row) in NHERF1 knockdown SKBR3 cells. All scale bars represent 10µM. C) PLA fluorescence showing protein-protein interactions between HER2/ Erbin, Erbin/NHERF1, or Erbin/Ezrin in control, NSC668394 treated-, and NHERF1 knock-down SKBR3 cells. All scale bars represent 10µM. (n = 3) D) Quan tification of PLA results was performed by measuring the fluorescent intensity of PLA signals at the cell surface. HER2/Erbin (control n = 14, NSC668394 n = 14, NHERF1KD n = 14), Erbin/NHERF1 (control n = 19, NSC668394 n = 19, NHERF1KD n = 19), Erbin/EZRIN (control n = 15, NSC668394 n = 15, NHERF1KD n = 15). Bar graphs represent the mean ± SEM. **** denotes p < 0.00005. (n = 3). E) A working model proposing how HER2 mediated disruption of apical/ basal polarity creates the opportunity for the formation of a multi-protein signaling complex that includes HER2, Erbin, PMCA2, NHERF1, and Ezrin. Cre ated with BioRender.com

Article Snippet: Erbin knockdown cell line A stable cell line expressing shRNA directed against Erbin was generated by transducing cells with commercially prepared lentiviruses containing three individual shRNA targeting Erbin mRNA (sc-40541-V) (Santa Cruz).

Techniques: Immunofluorescence, Knockdown, Fluorescence, Protein-Protein interactions, Control, Disruption