human skeletal muscle whole tissue lysate (adult whole normal)  (Bio-Techne corporation)

Journal: JCI insight

Article Title: A modifier screen identifies DNAJB6 as a cardiomyopathy susceptibility gene.

doi: 10.1172/jci.insight.88797

Figure Lengend Snippet: Figure 2. The GBT0411 mutant was tagged to the dnajb6b(L) isoform that predominantly expresses in the heart. (A) Northern blot analysis of dnajb6b(S) and dnajb6b(L) isoforms in indicated 3-month-old adult fish tissues demonstrated a predominant expression of dnajb6b(L) in the fish heart and skeletal muscle. (B) Western blot analysis (left) and quantification (right) of Dnajb6(S) and Dnajb6(L) isoforms expression in 1-year-old WT C57BL/6 mouse tissues indicated a predominant expression of Dnajb6(L) in the mouse heart. Values are presented relative to heart expression. Data represent mean ±SEM. (C) Western blot analysis (left) and quantification (right) of DNAJB6(L) and DNAJB6(S) protein expression in human heart and skeletal muscle lysates. Values are presented relative to heart expression. Data represent mean ±SEM. (D) Kaplan-Meier survival curves of indicated fish injected with a single bolus of 20 μg/gbm doxorubicin (DOX). Cardiomyocyte-specific overexpression of zebrafish dnajb6b(L) rescued DOX-induced fish death in the GBT0411 heterozygous mutant. *P < 0.05, log-rank test. Comparison of Tg(cmlc2:dnajb6b(L)-EGFP); GBT0411+/– vs. GBT0411+/–.

Article Snippet: Human heart and skeletal muscle tissue lysates were purchased from Novus Biologicals LLC.

Techniques: Mutagenesis, Northern Blot, Expressing, Western Blot, Injection, Over Expression, Comparison

Journal: Oncogene

Article Title: IGF2BP1 controls cell death and drug resistance in rhabdomyosarcomas by regulating translation of cIAP1.

doi: 10.1038/onc.2014.90

Figure Lengend Snippet: Figure 1. IGF2BP1 is overexpressed in RMS and drives cIAP1 expression. (a) RMS cell lines were analysed by western blot for IGF2BP1, cIAP1 and GAPDH protein expression compared with HSMMs. (b) Western blot of four human primary eRMS tumours for IGF2BP1, cIAP1 and GAPDH protein expression compared with fetal and adult human skeletal muscle. (c) Human RMS primary tumours and normal skeletal muscle formalin-fixed, paraffin-embedded (FFPE) sections were analysed by immunohistochemistry for IGF2BP1 and cIAP1 expression (brown staining) and representative images shown. (d) Intensity scoring of the sections in (c) was analysed 1 = mild, 2 = moderate and 3 = strong expression. Spearman's correlation P- value of 0.033 between IGF2BP1 and cIAP1 staining intensity (n = 8 RMS, 9 normal muscle). (e) RH36 cells were transfected with non-targeting siRNA (siC) or IGF2BP1 siRNA (siIGF2BP1) for 72h, and IGF2BP1, cIAP1 and GAPDH protein expression were analysed by western blot analysis. Bar graphs show densitometry of IGF2BP1 and cIAP1 protein levels normalised to GAPDH. (f) RH36 cells were transfected with siC or siIGF2BP1 for 72h, and cIAP1 mRNA levels were analysed by RT–qPCR, compared with GAPDH. Results are representative of at least three biological replicates. Mean ± s.e.m. Student's t-test: NS, nonsignificant, *Po0.05, **Po0.01 and ***Po0.001.

Article Snippet: Control human and fetal skeletal muscle whole lysates were purchased from Novus Biologicals (Oakville, ON, Canada).

Techniques: Expressing, Western Blot, Immunohistochemistry, Staining, Transfection, Quantitative RT-PCR