Figure - available from: The FEBS Journal
This content is subject to copyright. Terms and conditions apply.
![The C‐terminus of HSF2 binds to TLN1. (A) An in vitro fluorescent polarization assay between a fluorescently labeled mouse HSF2 peptide containing amino acids 519–535 and three regions of mouse TLN1 (R4–R8, R9–R12, R13–DD). The LD motif of KANK1 was used as a positive control (R1–R3, R4–R8, R9–R12, R13–DD). (B) A schematic overview of fusion proteins containing human HSF1 or HSF2 and monomeric Enhanced Green Fluorescent Protein (mEGFP). The C‐terminal amino acids of HSF1 (504–529) and HSF2 (511–536) were fused to mEGFP and named as HSF1‐c25 and HSF2‐c25. Mock contained only the mEGFP. (C) Immunoblot analysis of TLN1 and GFP in GFP‐Trap co‐IP and input samples from PC‐3 cells expressing Mock, HSF1‐c25, or HSF2‐c25 constructs. TLN1‐HSF2 interaction is indicated with an asterisk. (D) Immunoblot analysis of HSF2 expression in PC‐3 cells. PC‐3 cells were transfected with scrambled (Scr) or HSF2‐targeting shRNA constructs (shHSF2) [48]. Tubulin was used as a loading control. (E) Immunoblot analysis of TLN1 in HSF2 co‐IP samples from PC‐3 cells. IgG was used as a negative control. The cells were transfected as in (D). TLN1‐HSF2 interaction is indicated with an asterisk. The immunoblots in C–E are representative figures of two biological replicates. (F) Representative images of spheroid‐like structures in ultra‐low attachment plates, scale bar 200 μm (left panel). Quantitative analysis of spheroid area (right panel). The area was quantified in imagej. Results were plotted as independent data points with mean ± SEM. ****P‐value ≤ 0.0001. The data represents three biological replicates.](publication/384072702/figure/fig5/AS:11431281287968349@1730479753055/The-C-terminus-of-HSF2-binds-to-TLN1-A-An-in-vitro-fluorescent-polarization-assay.png)
The C‐terminus of HSF2 binds to TLN1. (A) An in vitro fluorescent polarization assay between a fluorescently labeled mouse HSF2 peptide containing amino acids 519–535 and three regions of mouse TLN1 (R4–R8, R9–R12, R13–DD). The LD motif of KANK1 was used as a positive control (R1–R3, R4–R8, R9–R12, R13–DD). (B) A schematic overview of fusion proteins containing human HSF1 or HSF2 and monomeric Enhanced Green Fluorescent Protein (mEGFP). The C‐terminal amino acids of HSF1 (504–529) and HSF2 (511–536) were fused to mEGFP and named as HSF1‐c25 and HSF2‐c25. Mock contained only the mEGFP. (C) Immunoblot analysis of TLN1 and GFP in GFP‐Trap co‐IP and input samples from PC‐3 cells expressing Mock, HSF1‐c25, or HSF2‐c25 constructs. TLN1‐HSF2 interaction is indicated with an asterisk. (D) Immunoblot analysis of HSF2 expression in PC‐3 cells. PC‐3 cells were transfected with scrambled (Scr) or HSF2‐targeting shRNA constructs (shHSF2) [48]. Tubulin was used as a loading control. (E) Immunoblot analysis of TLN1 in HSF2 co‐IP samples from PC‐3 cells. IgG was used as a negative control. The cells were transfected as in (D). TLN1‐HSF2 interaction is indicated with an asterisk. The immunoblots in C–E are representative figures of two biological replicates. (F) Representative images of spheroid‐like structures in ultra‐low attachment plates, scale bar 200 μm (left panel). Quantitative analysis of spheroid area (right panel). The area was quantified in imagej. Results were plotted as independent data points with mean ± SEM. ****P‐value ≤ 0.0001. The data represents three biological replicates.
Source publication
Heat shock factor 2 (HSF2) is a versatile transcription factor that regulates gene expression under stress conditions, during development, and in disease. Despite recent advances in characterizing HSF2‐dependent target genes, little is known about the protein networks associated with this transcription factor. In this study, we performed co‐immunop...
