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![Identification of the HSF2 interactome in mouse testes. (A) A schematic figure of the workflow. Isolated mouse testes were lysed, and endogenous HSF2 protein was immunoprecipitated with an antibody specific for HSF2. IgG was used as a negative control. The HSF2 co‐immunoprecipitated (co‐IP) proteins were subsequently identified through liquid chromatography–tandem mass spectrometry (LC–MS/MS). (B) Immunoblot analysis of HSF2 and HSF1 in corresponding co‐IP and input samples derived from mouse testes. HSF1 was used as a positive control and IgG as a negative control. The immunoblots are representative figures of two biological replicates run on different gels. (C) Venn diagram showing shared and distinct proteins identified by LC–MS/MS in the HSF2 co‐IP sample and the IgG negative control. After applying cut‐off criteria (peptide spectrum matches [PSMs] > 2 and a ratio of HSF2 PSMs/IgG PSMs > 3), a total of 105 proteins were identified. (D) Gene ontology (GO) analysis showing terms associated with the 105 HSF2‐binding partners. The molecular function GO terms were ranked according to their false discovery rate (FDR) and the redundant terms were withdrawn. The top five GO term categories are shown. (E) HSF2‐binding partners within the top five categories shown in D. (F) Immunoblot analysis of MSH2, SMC2, TOP2A, and TLN1 in HSF2 and HSF1 co‐IP and input samples derived from mouse testis. IgG was used as a negative control. The immunoblots are representative figures of three biological replicates run on different gels.](publication/384072702/figure/fig1/AS:11431281287998745@1730479750983/Identification-of-the-HSF2-interactome-in-mouse-testes-A-A-schematic-figure-of-the.png)
Identification of the HSF2 interactome in mouse testes. (A) A schematic figure of the workflow. Isolated mouse testes were lysed, and endogenous HSF2 protein was immunoprecipitated with an antibody specific for HSF2. IgG was used as a negative control. The HSF2 co‐immunoprecipitated (co‐IP) proteins were subsequently identified through liquid chromatography–tandem mass spectrometry (LC–MS/MS). (B) Immunoblot analysis of HSF2 and HSF1 in corresponding co‐IP and input samples derived from mouse testes. HSF1 was used as a positive control and IgG as a negative control. The immunoblots are representative figures of two biological replicates run on different gels. (C) Venn diagram showing shared and distinct proteins identified by LC–MS/MS in the HSF2 co‐IP sample and the IgG negative control. After applying cut‐off criteria (peptide spectrum matches [PSMs] > 2 and a ratio of HSF2 PSMs/IgG PSMs > 3), a total of 105 proteins were identified. (D) Gene ontology (GO) analysis showing terms associated with the 105 HSF2‐binding partners. The molecular function GO terms were ranked according to their false discovery rate (FDR) and the redundant terms were withdrawn. The top five GO term categories are shown. (E) HSF2‐binding partners within the top five categories shown in D. (F) Immunoblot analysis of MSH2, SMC2, TOP2A, and TLN1 in HSF2 and HSF1 co‐IP and input samples derived from mouse testis. IgG was used as a negative control. The immunoblots are representative figures of three biological replicates run on different gels.
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...
