Journal: Scientific Reports

Article Title: A fixation method for the optimisation of western blotting

doi: 10.1038/s41598-019-43039-3

Figure Lengend Snippet: Fixation-dependent differences in lectin staining intensities when using PVDF and nitrocellulose membranes. Pooled human serum proteins (3 μg) were separated on 10% SDS-PAGE, and the proteins were transferred onto PVDF and nitrocellulose membranes, the whole membrane was cutted into five pieces for subsequently fixation treatments and followed by staining with lectins (LCA and SNA). Lane i, CBB staining; lane ii, no fixation; lane iii, drying at room temperature; lane iv, sample heating at 100 °C; lane v, organic solvent (acetone and 50% methanol for PVDF and nitrocellulose membranes, respectively) treatments at room temperature; lane vi, organic solvent treatments followed by sample heating at 100 °C. All treatments were applied for 30 min. Left, WB pattern; right, quantitative analysis (n = 3 individual experiments). The exposure times were the same in all procedures. Band intensities were analysed and compared using Image Lab software (Bio-Rad Laboratories) and GraphPad Prism version 6. ** Significantly different p < 0.01, *** p < 0.001, **** p < 0.0001. All values are means ± S.E. (error bars).

Article Snippet: Biotinylated lectins, LCA, SNA, PHA-E, PHA-L, and AAL (Supplementary Table ), were purchased from Vector Laboratories Inc. (Burlingame, CA, USA).

Techniques: Staining, SDS Page, Software

Journal: PLoS Genetics

Article Title: The steroid hormone 20-hydroxyecdysone binds to dopamine receptor to repress lepidopteran insect feeding and promote pupation

doi: 10.1371/journal.pgen.1008331

Figure Lengend Snippet: A . Apoptosis signal in HaEpi cells after DMSO, 20E (5 μM), dsGFP +20E (5 μM), dsDopEcR+ 20E (5 μM) and DA (10 μM) +20E (5 μM) treatment by using the NucView caspase-3 activity assay kit. a . The ratio of apoptotic cells (green) to the total cells (blue) in the field view was obtained. B . Proliferation signal in HaEpi cells after DMSO, 20E (5 μM), dsGFP +20E (5 μM), dsDopEcR+ 20E (5 μM) and DA (10 μM) +20E (5 μM) treatment by using the 5-ethynyl-2′-deoxyuridine (EdU) kit (Ribobio, Guangzhou, China). b . The ratio of proliferation cells (green) to the total cells (blue) in the field view was obtained. DAPI stained the nucleus (blue). Statistical analysis using the data from 100 × 3 cells. The yellow bars represent 100 μM. C . Western blot analysis of 20E (5 μM) or DA (10 μM) induced proteins phosphorylation for 30 min. a. b . and c . AKT-RFP-His, AKT -RFP-His, and CDK10-RFP-His. 7.5% SDS-PAGE gel. Error bars show the mean ± SD of three times’ biological repetition. ImageJ software was used to transform the image data. Significant differences were calculated by Student’s t test (* p < 0.05; ** p < 0.01) or one-way analysis of variance (ANOVA, p < 0.05).

Article Snippet: The NucView 488 caspase-3 assay kit (NO. 30029 Biotium, Hayward, USA) was used to detect the activity of caspase-3 in the HaEpi cells, according to the manufacturer’s instructions.

Techniques: Caspase-3 Activity Assay, Staining, Western Blot, SDS Page, Software