Picture from Chen, Jian et al. MicroPubl Biol

Figure 1. Novel primer design strategy to detect single nucleotide polymorphisms (SNPs) by PCR.: (A) Schematic showing the primer design strategy to discriminate between the variant and wild type alleles through PCR (compare I and II, III and IV). The SNP residue is denoted as position 0. G, in red, is the variant allele, while T, in green, is the wild type allele. The forward PCR primer was designed to specifically detect either the variant or wild type allele, with the 3' end starting at the SNP residue (G or T at position 0). For the allele to be detected, there is no mismatch at position 0, while for the other allele, there is a mismatch (I versus II, and III versus IV). To increase the specificity for the allele to be detected by PCR, a second change (C to A in these examples, in blue) was introduced at the -2 position, two nucleotides upstream of the SNP position. For examples I and II, the variant forward primer can bind more efficiently to the variant allele (one mismatch in I) than the wild type allele (two mismatches in II), resulting in a more robust PCR amplification of the variant allele. Similarly for examples III and IV, the wild type forward primer can detect the wild type allele (shown in III) more effectively than the variant allele (shown in IV). "X" indicates mismatch,

Picture from Chen, Jian et al. (2020) MicroPubl Biol

Comparison of TIR1 transgene mediated degradation in C. elegans germline stem cells by qRT-PCR: (A) Schematic showing four distinct TIR1 transgenes driven by different promoters, indicated by the gene name followed by 'p', for promoter. mRuby and BFP are fluorescent proteins, F2A is a self-cleavable linker. Not shown is the substrate, LAG-1::degron. (B) Schematic illustrating sample collection for qRT-PCR analysis. The animals were dissected and the extruded gonads collected for RNA purification and then qRT-PCR analysis. (C & D) The relative mRNA level of LAG-1 dependent transcriptional targets lst-1 (C) and sygl-1 (D) from dissected gonad samples of the indicated genotype and treatment. q175 is a glp-1 null allele, where the mRNA level of sygl-1 and lst-1 were normalized by setting their levels to one for this genotype. ar202 is a glp-1 gain of function allele. Results are from three biological replicates, except for sun-1p::TIR1 (two replicates). Bar shows mean and standard deviation. Each dot represents a single data point. ns for not significant, * for p<0.01, and ** for p<0.001 by two-tailed t-test.

Picture from Chen P-J et al. (2000) Development "TRA-1A regulates transcription of fog-3, which controls germ cell fate in C. ...."

The period of high fog-3 expression coincides with spermatogenesis. Northern blot of N2 total RNA, isolated from different developmental stages. L1, first larval stage; yA, young adult; A, adult. The blot was probed with an anti-fog-3 probe (Chen et al., 2000) and then with a probe to the 26s ribosomal RNA, to test for loading errors.

Picture from Chen P-J et al. (2000) Development "TRA-1A regulates transcription of fog-3, which controls germ cell fate in C. ...."

Figure 2. The period of high fog-3 expression coincides with spermatogenesis. (A) Northern blot of N2 total RNA, isolated from different developmental stages. L1, first larval stage; yA, young adult; A, adult. The blot was probed with an anti-fog-3 probe (Chen et al., 2000) and then with a probe to the 26s ribosomal RNA, to test for loading errors. (B) Northern blot of him-5(e1490) total RNA, processed as described in A. These populations contain 30% males and 70% hermaphrodites (Hodgkin et al., 1979).

Picture from Larsen MK et al. (2006) Mol Biol Cell "MAA-1, a novel acyl-CoA-binding protein involved in endosomal vesicle ...."

Figure 3. MAA-1 localizes to endosomal and Golgi compartments. Photomicrographs of the intestines of worms viewed with confocal fluorescence microscopy. The five panels on the left show GFP fluorescence in worms expressing MAA-1::GFP fusion proteins. The five middle panels show staining of the same worms with markers for different membrane organelles. FM4-64 is a membrane-intercalating dye that labels early and late endosomes. LysoTracker labels late endosomes and lysosomes. An antibody that recognizes the ER retention signal HDEL was used to label the ER. An antibody that recognizes the Golgi protein GRASP55 was used to label Golgi. Staining with this antibody substantially overlaps that obtained with a marker for Golgi in C. elegans, alpha-mannosidase (Chen et al., 2006) (Supplemental Figure S2). Because the epitope from human GRASP55 used to raise the antibody is almost completely conserved in the worm orthologue, it is likely that the antibody recognizes worm GRASP55. RME-1 labels the ERC. The five panels on the right show merged images of the respective left and middle panels. Bar, 5 um. Selected images of worms at lower magnification are shown in Supplemental Figure S3.

Picture from Siete C et al. (2024) MicroPubl Biol "Immobilization of C. elegans with different concentrations of an ...."

Figure 1. Comparison of axonal transport of TIR-1:GFP in AWC axons of worms immobilized with different concentrations of tetramisole and polystyrene microbeads:(A) Representative kymographs of moving TIR-1::GFP in AWC axons of worms immobilized with 0.5-7.5 mM tetramisole (Tet) or microbeads. The line within two blue arrowheads represents a retrograde trafficking event toward the proximal axon (P) or the ­­­­­AWC cell body; the line between two red arrowheads represents an anterograde trafficking event toward the distal (D) axon. (B) The percentage of anterograde and retrograde trafficking events. The average number of trafficking events per animal imaged is 19 (0.5 mM Tet), 24 (1 mM Tet), 21 (2 mM Tet), 19 (7.5 mM Tet), and 13 (microbeads). Z-test was used to determine p-values. Error bars, SE of proportion. ns, not significant. (C) The average velocity of anterograde and retrograde trafficking events. Red and blue lines indicate the comparison of anterograde and retrograde velocities, respectively. Student's t-test was used to determine p-values. Error bars, SEM. ns, not significant. The significance level 0.01 was adjusted to 0.0025 (0.01/4) using the Bonferroni method (Chen, Feng, & Yi, 2017) in (B) and (C).