Journal: eLife

Article Title: insomniac links the development and function of a sleep-regulatory circuit

doi: 10.7554/eLife.65437

Figure Lengend Snippet:

Article Snippet: Samples were incubated overnight at 4°C in rat anti-HA (1:100; Sigma, 11867431001) and mouse anti-Brp (1:20, DSHB, nc82) antibodies prepared in 5% NDS in PBST.

Techniques:

Journal: eLife

Article Title: Fruitless mutant male mosquitoes gain attraction to human odor

doi: 10.7554/eLife.63982

Figure Lengend Snippet: ( A, B ) Arm-next-to-cage assay schematic A and image B with female (top) and male (bottom) Aedes aegypti mosquitoes. ( C ) Percent mosquitoes next to arm measured every 30 s. Data are mean ± s.e.m., n = 6 trials, n = 20 mosquitoes/trial; *p<0.05, Mann-Whitney test for each time point. ( D, E ) Schematic of Aedes aegypti fruitless genomic locus D and sex-specific splicing region with RNA-seq read evidence E . ( F ) Phylogeny of mosquito species with outgroup Drosophila melanogaster , with conserved fruitless exon structure inferred from de novo transcriptome assembly. In E , F , coding and non-coding exons are represented by filled and open dashed bars, respectively. Toxorhynchites rutilus and Culex salinarius images were used to represent Toxorhynchites amboinensis or Culex quinquefasciatus , respectively. See acknowledgments for photo credits. Cartoons indicate blood-feeding (blood drop) and non-blood-feeding (flower) species. ( G, H ) Aedes aegypti fruitless exon usage based on male and female RNA-seq data (normalized counts) from the indicated tissue plotted for each exon G and m, f, and c1 exons H n = 3–4 independent RNA-seq replicates . ( I ) Schematic of generation of fruitless ∆M and fruitless ∆M-tdTomato mutants. Figure 1—source data 1. Source data for .

Article Snippet: Brains were incubated in 0.25% PBT plus 2% normal goat serum with primary antibodies at the following dilutions: rabbit anti-RFP (Rockland 600-401-379) 1:200 and mouse anti- Drosophila melanogaster Brp (nc82) 1:5000.

Techniques: MANN-WHITNEY, RNA Sequencing

Journal: eLife

Article Title: Fruitless mutant male mosquitoes gain attraction to human odor

doi: 10.7554/eLife.63982

Figure Lengend Snippet: ( A ) Heat-seeking assay schematic. A 20 s pulse of 10% CO 2 is added to the assay. ( B ) Percent of animals on Peltier. Data are mean ± s.e.m., n = 6 trials/temperature, n = 50 mosquitoes/trial. Data labeled with different letters are significantly different from each other, within each temperature. ( C ) Schematic of human odor host-seeking assay (left) and stimuli (right). ( D ) Percent of attracted animals. Data are mean ± s.e.m., n = 8–14 trials/group, n = 17–28 mosquitoes/trial. ( E–F ) Summary of results and model of gain of fruitless function in Aedes aegypti . Photo credit: Aedes aegypti (Alex Wild); D. melanogaster (Nicolas Gompel). In B , D , data labeled with different letters are significantly different from each other (Kruskal-Wallis test with Dunn’s multiple comparisons, p<0.05). In B , comparisons are made between genotypes at each temperature. In D , comparisons are made across all genotypes and stimuli. Figure 5—source data 1. Source data for .

Article Snippet: Brains were incubated in 0.25% PBT plus 2% normal goat serum with primary antibodies at the following dilutions: rabbit anti-RFP (Rockland 600-401-379) 1:200 and mouse anti- Drosophila melanogaster Brp (nc82) 1:5000.

Techniques: Labeling

Journal: Cells

Article Title: Geniposide Ameliorated Dexamethasone-Induced Cholesterol Accumulation in Osteoblasts by Mediating the GLP-1R/ABCA1 Axis

doi: 10.3390/cells10123424

Figure Lengend Snippet: GEN ameliorated DEX-induced cholesterol accumulation by increasing ABCA1 expression. ( A , B ) The in vivo immunohistochemical examination of ABCA1 expression was conducted. ( C – E ) The protein expression of ABCA1 and ApoA-I was detected by western blot in DEX-treated MC3T3-E1 cells. ( F ) The level of the total intracellular cholesterol was measured using the ELISA kit in MC3T3-E1 cells co-treated with DEX and DIDS (200 μM). ( G – I ) The protein expression of ABCA1 and ApoA-I was detected by western blot in DEX/DIDS-treated MC3T3-E1 cells. ( J ) The ALP staining and the Alizarin Red S staining assays were conducted (×100 magnification). ( K – M ) The protein expression of RUNX2 and OPN was detected by western blot in DEX/DIDS-treated MC3T3-E1 cells. All experiments were implemented separately in triplicate. * p < 0.05; ** p < 0.01. OIM, osteogenic induction medium. NC, negative control; 50 mg/kg, Dex + 50 mg/kg GEN; 100 mg/kg, Dex + 100 mg/kg GEN.

Article Snippet: After being blocked in TBS containing 5% skimmed milk for 1 h, the membranes were co-incubated with the primary antibodies at 4 °C overnight against RUNX2 (1:1000 dilution; MyBioSource, cat.no.127554, San Diego, CA, USA), OPN (1:1000 dilution; Proteintech, cat.no.22952-1-AP, Rosemont, IL, USA), GLP-1R (1:1000 dilution; ABGENT, cat.no.AP52040), ABCA1 (1:1000 dilution; Affinity, cat.no.DF8233), apoA-I (1:1000 dilution; Affinity, cat.no.DF6264), and GAPDH (1:1000 dilution; Affinity, cat.no.AF7021), and then with HRP-labeled goat anti-rabbit secondary antibody (1:5000 dilution; Boster Biological Technology, Wuhan, China).

Techniques: Expressing, In Vivo, Immunohistochemical staining, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Negative Control

Journal: Cells

Article Title: Geniposide Ameliorated Dexamethasone-Induced Cholesterol Accumulation in Osteoblasts by Mediating the GLP-1R/ABCA1 Axis

doi: 10.3390/cells10123424

Figure Lengend Snippet: GEN promoted ABCA1-mediated cholesterol metabolism in a GLP-1R-dependent manner. ( A , B ) The in vivo immunohistochemical examination of GLP-1R expression was conducted. ( C , D ) The protein expression of GLP-1R was determined by western blot in DEX-treated MC3T3-E1 cells. ( E ) The ALP staining and the Alizarin Red S staining assay were performed in EX-treated MC3T3-E1 cells (×100 magnification). ( F ) The level of the total intracellular cholesterol was measured using the ELISA kit. ( G – K ) The protein expression of GLP-1R, ABCA1, RUNX2, and OPN was detected by western blot. All experiments were implemented separately in triplicate. * p < 0.05; ** p < 0.01. NC, negative control; 50 mg/kg, Dex + 50 mg/kg GEN; 100 mg/kg, Dex + 100 mg/kg GEN; EX, Exendin9-39.

Article Snippet: After being blocked in TBS containing 5% skimmed milk for 1 h, the membranes were co-incubated with the primary antibodies at 4 °C overnight against RUNX2 (1:1000 dilution; MyBioSource, cat.no.127554, San Diego, CA, USA), OPN (1:1000 dilution; Proteintech, cat.no.22952-1-AP, Rosemont, IL, USA), GLP-1R (1:1000 dilution; ABGENT, cat.no.AP52040), ABCA1 (1:1000 dilution; Affinity, cat.no.DF8233), apoA-I (1:1000 dilution; Affinity, cat.no.DF6264), and GAPDH (1:1000 dilution; Affinity, cat.no.AF7021), and then with HRP-labeled goat anti-rabbit secondary antibody (1:5000 dilution; Boster Biological Technology, Wuhan, China).

Techniques: In Vivo, Immunohistochemical staining, Expressing, Western Blot, Staining, Enzyme-linked Immunosorbent Assay, Negative Control

Journal: EMBO Reports

Article Title: Pumilio differentially binds to mRNA 3′ UTR isoforms to regulate localization of synaptic proteins

doi: 10.1038/s44319-025-00401-z

Figure Lengend Snippet: Reagents and tools table

Article Snippet: For NMJ immunofluorescence, mouse anti-Syt1 (DSHB #3H2 2D7-s), mouse anti-BRP (DSHB #nc82-c), goat anti-HRP (Jackson ImmunoResearch #123-605-021) and donkey anti-mouse (Abcam #ab150106) were used at concentrations: 1:50, 1:50, 1:50, and 1:500, respectively.

Techniques: Recombinant, Sequencing, Magnetic Beads, SYBR Green Assay, cDNA Synthesis, Protease Inhibitor, Plasmid Preparation, Software, Membrane, Microscopy