Journal: PLoS ONE

Article Title: The Mechanisms Underlying α-Amanitin Resistance in Drosophila melanogaster : A Microarray Analysis

doi: 10.1371/journal.pone.0093489

Figure Lengend Snippet: Single gene analysis for Ama-KTT/M/2 versus Canton-S on no toxin (group 2 versus 1).

Article Snippet: The primer pairs used were a part of the Taqman Gene Expression Assays kit (Applied Biosystems): Dm02361072_s1, Dm01831596_g1, Dm01840671_g1, Dm01830394_g1, Dm01822311_g1, Dm01804633_g1, Dm01799869_s1, Dm02147253_g1, Dm01817955_g1, Dm02152265_s1, Dm01826948_s1, and Dm02374415_g1.

Techniques:

Journal: Skeletal muscle

Article Title: Myostatin as a mediator of sarcopenia versus homeostatic regulator of muscle mass: insights using a new mass spectrometry-based assay.

doi: 10.1186/s13395-015-0047-5

Figure Lengend Snippet: Fig. 1 Calibration curves for LC-MS/MS measures of myostatin, propeptide, GASP-1, and FLRG. Linear calibration curves for recombinant intact myostatin (a), propeptide (b), GASP-1 (c), and FLRG (d) diluted into bovine serum albumin over a concentration range of 0.031 to 1.00 nM

Article Snippet: Recombinant human GASP-1 (cat# 2070-GS- 025), recombinant human FLRG (cat# 1288-F3-025), biotinylated anti-myostatin antibody (cat# BAF788), biotinylated anti-GASP1 antibody (cat# BAF2070), and biotinylated anti-FLRG antibody (cat# BAF1288) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Liquid Chromatography with Mass Spectroscopy, Recombinant, Concentration Assay

Journal: Skeletal muscle

Article Title: Myostatin as a mediator of sarcopenia versus homeostatic regulator of muscle mass: insights using a new mass spectrometry-based assay.

doi: 10.1186/s13395-015-0047-5

Figure Lengend Snippet: Fig. 3 Circulating FLRG and GASP-1 concentrations by LC-MS/MS in younger and older women and men. Box plots (25–75 percentile) and whiskers (Tukey method) showing comparisons of a FLRG, b GASP-1, c FLRG relative to myostatin, and d GASP-1 relative to myostatin between younger women (YW), older women (OW) and sarcopenic older women (SOW). Comparisons of myostatin inhibitors and their ratios to myostatin between corresponding groups of men are also illustrated (e-h). *, **, and *** denote p < 0.05, 0.01, and 0.001, respectively, compared to the younger group except when denoted with a bracket

Article Snippet: Recombinant human GASP-1 (cat# 2070-GS- 025), recombinant human FLRG (cat# 1288-F3-025), biotinylated anti-myostatin antibody (cat# BAF788), biotinylated anti-GASP1 antibody (cat# BAF2070), and biotinylated anti-FLRG antibody (cat# BAF1288) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Liquid Chromatography with Mass Spectroscopy

Journal: Skeletal muscle

Article Title: Myostatin as a mediator of sarcopenia versus homeostatic regulator of muscle mass: insights using a new mass spectrometry-based assay.

doi: 10.1186/s13395-015-0047-5

Figure Lengend Snippet: Fig. 5 Circulating FLRG and GASP-1 concentrations in women compared to men. Box plots (25–75 percentile) and whiskers (Tukey method) showing comparisons of a FLRG, b GASP-1, c FLRG relative to myostatin, and d GASP-1 relative to myostatin between younger women (YW) and younger men (YM) (top panel), older women (OW) and older men (OM) (middle panel), and sarcopenic OW (SOW) and sarcopenic OM (SOM) (bottom panel). *** denotes p < 0.001

Article Snippet: Recombinant human GASP-1 (cat# 2070-GS- 025), recombinant human FLRG (cat# 1288-F3-025), biotinylated anti-myostatin antibody (cat# BAF788), biotinylated anti-GASP1 antibody (cat# BAF2070), and biotinylated anti-FLRG antibody (cat# BAF1288) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques:

Journal: PLoS ONE

Article Title: Drug-Encoded Biomarkers for Monitoring Biological Therapies

doi: 10.1371/journal.pone.0137573

Figure Lengend Snippet: Glucuronidase assay substrates.

Article Snippet: Carboxyumbelliferyl β-D-glucuronide , CUGlcU , 386 nm , 445 nm , Marker Gene Technologies Inc., Eugene, USA.

Techniques: Marker

Journal: Investigative Ophthalmology & Visual Science

Article Title: Proteopathy Linked to Exon-Skipping Isoform of RGR-Opsin Contributes to the Pathogenesis of Age-Related Macular Degeneration

doi: 10.1167/iovs.64.13.41

Figure Lengend Snippet: Primers Used for PCR

Article Snippet: Lentivirus carrying the human RGR and RGR-d gene (Lv-RGR and Lv-RGR-d) was constructed commercially by Genechem Technology (Shanghai, China).

Techniques: Reverse Transcription

Journal: Investigative Ophthalmology & Visual Science

Article Title: Proteopathy Linked to Exon-Skipping Isoform of RGR-Opsin Contributes to the Pathogenesis of Age-Related Macular Degeneration

doi: 10.1167/iovs.64.13.41

Figure Lengend Snippet: Degradation of RGR-d protein by the ubiquitin-proteasome system. ARPE-19 cells were stably transfected by NC-, RGR-, or RGR-d–overexpressing lentivirus, referred to as NC, RGR, and RGR-d cells. ( A ) Quantitative PCR detected significantly increased expression mRNA levels of the RGR and RGR-d gene in the corresponding transfected ARPE-19 cells. ( B ) Western blotting detected Flag-tagged RGR and RGR-d protein levels under treatment of 4 µM MG132, a 26S proteasome inhibitor. β-Actin levels serve as a protein loading control. ( C ) Quantification of Western blotting results in B . ( D ) Immunofluorescence assay showing expression of RGR and RGR-d. RGR-d protein could only be detected in the presence of MG132. Scale bars : 25 µm. ( E ) Co-IP using precoated Flag-beads demonstrated the level of polyubiquitin chains on RGR-d and RGR protein using the anti-HA antibody. The Myc-beads served as a negative control to exclude nonspecific binding by the Flag-beads. β-Actin levels serve as a protein loading control for input whole-cell lysates. Values in A and C are the mean ± SD. Statistics: unpaired two-tailed t -test ( A ), one-way ANOVA followed by Fisher's least significant difference (LSD) post hoc test ( C ). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Lentivirus carrying the human RGR and RGR-d gene (Lv-RGR and Lv-RGR-d) was constructed commercially by Genechem Technology (Shanghai, China).

Techniques: Ubiquitin Proteomics, Stable Transfection, Transfection, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Control, Immunofluorescence, Co-Immunoprecipitation Assay, Negative Control, Binding Assay, Two Tailed Test

Journal: Investigative Ophthalmology & Visual Science

Article Title: Proteopathy Linked to Exon-Skipping Isoform of RGR-Opsin Contributes to the Pathogenesis of Age-Related Macular Degeneration

doi: 10.1167/iovs.64.13.41

Figure Lengend Snippet: In vitro and in vivo analyses of cytotoxicity caused by RGR-d accumulation. Cytotoxicity caused by RGR-d accumulation. ARPE-19 cells were transfected by NC-, RGR-, or RGR-d–overexpressing lentivirus. ( A ) Growth curve showed that the proliferation of RGR-d cells slightly slowed down under normal culture. ( B ) After treatment of 2 µM MG132, the RGR-d cells exhibited the most severe cell morphology changes. ( C ) Cell viability assay by CCK-8 analysis showed the most extensive impaired cell viability of RGR-d cells when treated with different concentrations of MG132. Values are described as percentages to DMSO-treated NC cells, mean ± SD. ( D ) ZO-1 images show severely damaged RPE integrity in RGR-d mice compared with wild-type mice at 23 months old. ( E ) TUNEL assay of fundus sections showing the photoreceptor apoptotic level in peripheral and central fundus regions of RGR-d mice aged 23 months. Scale bars : 200 µm ( B ), 20 µm ( D ), and 100 µm ( E ). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Lentivirus carrying the human RGR and RGR-d gene (Lv-RGR and Lv-RGR-d) was constructed commercially by Genechem Technology (Shanghai, China).

Techniques: In Vitro, In Vivo, Transfection, Viability Assay, CCK-8 Assay, TUNEL Assay