EMBO/soda-nel huggingface dataset:
HACE1-dependent protein degradation
            provides cardiac protection in response to haemodynamic stress (2014)
f7

(a,b) Hace1−/−
MEF cells were stably transfected with HA-tagged WT HACE1 (HA-HACE1) and empty vector MSCV.
(a) Immunoblot for HA shows stable expression of HA-tagged
HACE1 in
HA-HACE1 MEF cell
lysate. (b) HA-HACE1 MEF and control MSCV MEF cell lysates were
subjected to Co-IP with anti-HA antibody and coomassie blue staining shows a
couple of distinct bands only detected in HA-HACE1 MEF IP pull-down as indicated
by red arrows. (c–f) Identification of
HACE1 interacting
proteins by MS analysis of Co-IP products. (c) Hierarchical
clustering (heat) map demonstrating the clustering of proteins based on
their expression intensities in HA-HACE1 MEF and control MSCV MEF HA-IP pull-down products.
To the left of the heat map, selected annotations found to be enriched in
the protein clusters are shown. n=5 for HA-HACE1 group and n=4 for
control MSCV group. (d) Pie chart shows enriched representative
HACE1-interacting
proteins according to molecular function by GO term. (e) Selected
HACE1-interacting
proteins identified by HA-IP MS were evaluated by western blot with the
indicated antibodies (IB). HA-HACE1 denotes HA-IP pull-down from HA-HACE1 MEF; MSCV denotes HA-IP
pull-down from control MSCV MEF; and input indicates HA-HACE1 MEF lysate before IP.
(f) HA-HACE1
MEF and control MSCV MEF lysates were subjected to co-IP with an
anti-Unc-45b antibody
and the pull-down were analysed by western blot with an anti-HA antibody.
(g,h) HA-IP pull-down products from HA-HACE1 MEF (HA-HACE1) and Hace1 KO MSCV MEF (MSCV)
immortalized in HA beads were further subjected to Co-Co-IP with WT mouse
heart lysates, and the pull-down products were analysed by western blot
using the indicated antibodies. HACE1 and those proteins with which it interacts were
still detectable (g). There were also increased ubiquitinated
proteins in the Co-Co-IP heart tissues (h).
HACE1-dependent protein degradation
            provides cardiac protection in response to haemodynamic stress (2014)
f8

(a–c) Representative western blots and (d)
quantification show increased accumulation of some of the HACE1-interacting proteins in the
Hace1−/−
sTAC myocardium. n=3 mice per group, error bars represent s.e.m.,
P-value between KO and WT sTAC as indicated (one-way analysis of
variance). Gapdh was used as a loading control.