Skip to main content
. Author manuscript; available in PMC: 2022 Dec 2.
Published in final edited form as: Nat Biomed Eng. 2021 Dec 2;5(12):1500–1516. doi: 10.1038/s41551-021-00823-9

Fig. 8: LINC complex disruption alters H3K9me3 peripheral enrichment and actin colocalization.

Fig. 8:

a) Illustration of adenoviral vectors for LINC complex disruption. The decoupling vector (K3) expressed a truncated nesprin-3 composed of the transmembrane (TM) and the KASH domain tagged with mNeptune2.5. The control vector (CTL) lacked the KASH domain necessary for LINC complexes integration. b) CMs were infected with either vector on day one and stained for H3K9me3 (shown) or H3K27me3 (Extended Fig. 11) on day two or day four. The K3 construct integrated successfully into the outer nuclear membrane of infected CMs (mNep2.5) while no distinct localization was observed for the control vector. Scale=5 µm. c) Decoupled cells (K3) showed abolished enrichment of overall and H3K9me3-marked chromatin compared to infected control cells (CTL). SEM; n=35 from 3 exp.; T-test: * p<0.05, *** p<0.001. d) Gene expression analysis of decoupled (K3) or non-infected control cells (NIC) compared to infected control cells (CTL). Expression of structural, but not transcriptional, cardiac genes was reduced in decoupled cells. SD; n=4; T-test: * p<0.05, ** p<0.01. e) CMs on soft (13kPa) PDMS were infected with K3 or CTL on day one and stained for H3K9me3 or H3K27me3 on day four. Colocalization of H3K9me3 with α-actinin 2 containing Z-disks (Actn2) was abrogated after LINC complex disruption while colocalization of H3K27me3 was increased. n=18 from 3 exp.; T-test: * p<0.05, *** p<0.001.