Journal: PLoS Genetics

Article Title: HSPB7 prevents cardiac conduction system defect through maintaining intercalated disc integrity

doi: 10.1371/journal.pgen.1006984

Figure Lengend Snippet: (A) Immunoblot analysis of the cardiac muscle showing HSPB7 constitutive expression from the embryonic stages to adulthood (E14.5 to P28) with multiple forms of HSPB7 at different molecular masses (arrows). (B) Subcellular localization of HSPB7 in the cardiac muscle of adult mice. The heart sections were stained with antibodies against HSPB7 (red) and desmoplakin (desmosome), α-actinin (Z-line), myomesin (M-line), N-cadherin (adhering junction), connexin 43 (gap junction), and cardiac-actin (I-bend). HSPB7 mainly localizes at the intercalated discs and is adjacent to the Z-line with a striated pattern. (C) Colocalization of HSPB7 with N-cadherin during development. Heart sections from the embryonic stages to adulthood (E14.5 to P56) were stained with antibodies against HSPB7 (red) and N-cadherin (green). The nucleus was visualized through Hoechst 33342 staining. Insets show the representative areas with higher magnification. Scale bar: 10 μm.

Article Snippet: The primary antibodies used included mouse monoclonal antibodies anti-α-actinin (clone EA-53, Sigma-Aldrich; 1:200.), anti-Actin, cardiac (clone AC1-20.4.2, Sigma-Aldrich; 1:200), anti-connexin43 (clone CXN-6, Sigma-Aldrich; 1:200), antidesmoplakin (clone DP2.15, Millipore Corporation; 1:200), anti-mMaC myomesin B4 (DSHB; 1:200), and anti-Vinculin (clone hVIN-1, Sigma-Aldrich); rabbit polyclonal antibodies anti-FLAG (F7425, Sigma-Aldrich; 1:200), anti-pan-cadherin (#C3678, Sigma-Aldrich; 1:200); goat polyclonal antibodies anti-FLNC (K-18, Santa Cruz Biotechnology; 1:100); and guinea pig polyclonal antibody anti-HSPB7 (G11W, LTK BioLaboratories; 1:200).

Techniques: Western Blot, Expressing, Staining

Journal: American journal of physiology. Cell physiology

Article Title: HSP90 modulates the myosin replacement rate in myofibrils.

doi: 10.1152/ajpcell.00245.2017

Figure Lengend Snippet: Fig. 2. Inhibition of heat shock protein 90 (HSP90) activity slows the myosin replacement rate in myofibrils. A–L: sarcomere structures were not disrupted by geldanamycin (GM) treatment. DMSO (A–C and G–I) or 5 g/ml GM (D–F and J–L)-treated cells were stained with indicated antibodies. C, F, I, and L are merged images of A and B, D and E, G and H, and J and K. Arrowheads depict myomesin-positive M-lines in the A-bands in A–F. All images were taken with a confocal microscope. Scale bars 10 m. M–O: relative fluo- rescence intensity after bleaching of green fluorescent protein-tagged myosin heavy chain 3 (GFP-MYH3) was measured in myotubes treated with DMSO or GM. Fluorescence signals were measured at indicated time points (M). Mobile fractions (%) of GFP-MYH3 were 47.0 5.9 in DMSO and 9.6 1.2 in GM (N). Half- lives (hours) of GFP-MYH3 were 6.7 1.2 in DMSO and 3.4 1.0 in GM (O). Values represent the mean SE. *P 0.05, significant difference compared with DMSO control. DMSO control myotubes, n 7; GM-treated myotubes, n 4.

Article Snippet: The antibodies used in this study were mouse anti-myomesin antibody (1:10 dilution; clone B4; Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA) (19), mouse anti-HSP90 antibody (1:400 dilution; clone D7 ; StressMarq Biosciences), and rabbit polyclonal anti-MYH antibody (1:500 dilution; kindly gifted by the late Prof. Howard Holtzer, University of Pennsylvania, Philadelphia).

Techniques: Inhibition, Activity Assay, Staining, Microscopy, Fluorescence, Control