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. 2021 May 16;13(6):e13074. doi: 10.15252/emmm.202013074

Figure 2. PLN expression profile and response to isoprenaline.

Figure 2

  • A
    Western immunoblot analysis of total PLN in non‐failing heart tissue (NFH), PLNic, and PLN p.Arg14del EHTs. Pentameric (PM) and monomeric (MM) form of PLN under boiled (B) and non‐boiled (NB) conditions.
  • B
    PLN protein quantification of PM and MM forms under non‐boiled condition. NFH (n = 5 protein samples from two different donor hearts), PLNic (n = 8, each replicate consists of a pool of 3 EHTs from three different separate batches), and PLN p.Arg14del (n = 8, each replicate consists of a pool of 3–4 EHTs from three different separate batches). Loading was normalized to α‐actinin. One‐way ANOVA of PM and MM forms with Tukey’s post‐test, mean ± SEM, * P < 0.05.
  • C
    Western immunoblot analysis of PLN pSer16 MM and PM in PLNic and p.Arg14del EHTs (non‐boiled) in the absence and presence of isoprenaline (ISO; 100 nM, loading was normalized to α‐actinin).
  • D
    PLN pSer16 quantification of PM and MM form (non‐boiled; loading was normalized to α‐actinin). PLNic ± isoprenaline (n = 8, each replicate consists of a pool of 3–4 EHTs from three different separate batches) and PLN p.Arg14del ± isoprenaline (n = 8, each replicate consists of a pool of 3–4 EHTs from three different separate batches). One‐way ANOVA of PM and MM forms with Sidak’s post‐test, *P < 0.05. Mean ± SEM.
  • E, F
    Force (E) and relaxation time (RT) (F) of PLNic and PLN p.Arg14del EHTs in Tyrode's solution (Ca2+ 1.8 mM), Tyrode’s solution (EC50 [Ca2+]: PLNic: 0.4–0.6 mM, PLN p.Arg14del: 0.7–0.8 mM), in the presence of ISO (100 nM) or carbachol (10 µM); PLNic (n = 27 EHTs from 4 batches), PLN p.Arg14del EHTs (n = 19 EHTs from 3 batches), mean ± SEM, one‐way ANOVA for PLNic or PLN p.Arg14del EHTs with Tukey’s post hoc test, *P < 0.05. The data are representative of n = 6 independent experiments.