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Journal: The Journal of Biological Chemistry
Article Title: HSP-1-specific nanobodies alter chaperone function in vitro and in vivo
doi: 10.1016/j.jbc.2026.111238
Figure Lengend Snippet: B12 and H5 detect HSP-1 from C. elegans’ lysate. A and B , western blots of day 2 C. elegans that were grown on control ( pos-1 RNAi) plates and then transferred to either pos-1 or hsp-1 RNAi plates for 24 h as day 1 adults. pos-1 encodes a zinc-finger transcription factor required for embryonic development and sterilizes worms without affecting hatched animals ( , , , , , ). Membranes were probed with either biotinylated ( A ) B12 or ( B ) H5 and α-tubulin (DSHB, Cat #12G10). C , western blots from immunoprecipitation assay using recombinant H5 or B12 as the primary antigen-binding agent and probed with an anti-HSC70 antibody (Proteintech, Cat #10654-1-AP) and α-tubulin (DSHB, Cat #12G10). D and E , example ( D ) western blot and ( E ) quantification of nanobody B12 expression (MTX298 ( hsp-16.48p::B12::HA )) induced by a 30-min heat shock at 37 °C (HA, Cell Signaling (C29F4) and α-tubulin (DSHB, Cat #12G10)). F and G , example ( F ) western blot and ( G ) quantification of nanobody B12 expression (MTX298 ( hsp-16.48p::B12::HA )) placed at 25 °C for indicated amount of time or induced by 30-min heat shock at 37 °C (HA, Cell Signaling (C29F4) and α-tubulin (DSHB, Cat #12G10). Induction was started in day 1 adults. H , co-immunoprecipitation assay of WT and MTX298 (“B12”) grown on indicated siRNA and collected 5 h post 30-min heat shock at 37 °C using magnetic anti-HA beads. Ordinary one-way ANOVA. p < 0.05 is considered statistically significant. HSP, heat shock protein; HA, hemagglutinin; HSC70, heat shock cognate 70.
Article Snippet: C , western blots from immunoprecipitation assay using
Techniques: Western Blot, Control, Immunoprecipitation, Recombinant, Binding Assay, Expressing, Co-Immunoprecipitation Assay