Journal: Autophagy

Article Title: Identification of autophagy as a longevity-assurance mechanism in the aging model Podospora anserina

doi: 10.4161/auto.28148

Figure Lengend Snippet: Figure 3. Characterization of the Paatg1∆ strain. ( A ) The germination rate of ascospores from perithecia of fertilized Paatg1 Δ (n = 10) and WT cultures (n = 10). P values ( P < 0.001) were determined in comparison with the wild-type sample by 2-tailed Wilcoxon rank-sum test. ( B ) Determination of the genotype ( Ble R resistance) of ascospores of a Paatg1 Δ × WT cross (n = 10). Colonies grown from germinated ascospores were transferred to phleomycin containing BMM medium. Grey/bright = spores germinated ( Ble S ); red/dark = Ble R spores germinated with Paatg1 deletion background. ( C ) Ascospore phenotype of spores from WT × WT and WT × Paatg1 Δ crosses. Scale bar: 20 µm. ( D ) Growth rates of the WT (n = 27) and the Paatg1 deletion strain (n = 27; P < 0.001). P values were determined in comparison with the wild-type sample by 2-tailed Wilcoxon rank-sum test. ( E ) Life span of monokaryotic wild-type (n = 27; median life span = ~25 d) and Paatg1 Δ (n = 27; median life span = ~21 d; P < 0.001) isolates on M2 medium at 27 °C. P values were determined in comparison with the wild-type sample by 2-tailed Wilcoxon rank-sum test. ( F ) Western blot analysis of total protein extracts from the Paatg1 deletion mutant and WT strains grown on CM or CM-N media, respectively. Incubation with anti-PaPRE3 (corresponding to the β1 subunit of the 20S proteasome) and anti-PaGLO1 antibody (glyoxalase 1) showed different protein amounts in the WT and Paatg1 Δ strain and in response to nitrogen starvation compared with anti-SOD1 (loading control). ( G ) Life span of monokaryotic wild-type (n = 11; median life span = ~80 d) and Paatg1 Δ (n = 20; median life span = ~25 d; P < 0.001) isolates at 27 °C on M2-N medium. P values were determined in comparison with the wild-type sample by 2-tailed Wilcoxon rank-sum test. ( H ) Growth rates of the WT (n = 11) and the Paatg1 deletion strain (n = 20; P < 0.01) on M2-N medium at 27 °C. P values were determined in comparison with the wild-type sample by 2-tailed Wilcoxon rank-sum test. M2 medium: supplemented with nitrogen (0.5 g/L urea); N: M2-medium without nitrogen.

Article Snippet: The following primary antibodies were used: Anti-GFP (mouse, 1:10000 dilution, Sigma-Aldrich, G6795), anti-PaGLO1 (1: 2000 dilution, NEP, Frankfurt, Germany) raised against a specific peptide ([AC]-CVQNERFADKANF-[OH]) of PaGLO1 (glyoxalase1) previously described in Scheckhuber et al., anti-SOD1 (1:2000 dilution) from Biomol Stressgen (Hamburg, Germany) (SOD100) and Anti-PaPRE3 (1:2500 dilution) raised against a specific synthetic peptide ([H]-LYLPDTDYKVRHEN-[OH]; Sigma) of PaPRE3 (corresponding to the β1 subunit of the 26S proteasome).

Techniques: Western Blot, Mutagenesis, Incubation

Journal: iScience

Article Title: Venom exaptation and adaptation during the trophic switch to blood-feeding by kissing bugs

doi: 10.1016/j.isci.2024.110723

Figure Lengend Snippet:

Article Snippet: HEK293 cells stably expressing human Na V 1.7 co-expressed with the β1 auxiliary subunit , SB Drug Discovery , N/A.

Techniques: Virus, Recombinant, Sequencing, Clinical Proteomics, Stable Transfection, Expressing, Drug discovery, Plasmid Preparation, Software, Coagulation, Inverted Microscopy