Picture from Barbosa DJ et al. (2017) PLoS Genet "Dynactin binding to tyrosinated microtubules promotes centrosome centration ...."

S2 Fig. Dynein targeting to MT tips depends on dynactin.(A) Cortical confocal section of one-cell embryos in metaphase co-expressingendogenous dynein heavy chainDHC-1::GFP and EBP-2::mKate2, showing that depletionof p150DNC-1delocalizes dynein from MT tips. Images are maximum intensity projectionsover time (12 images acquired every 5 s). Scale bar, 5 um; insets, 2 um.(B) Quantification of dynein heavy chainDHC-1::GFP levels at MT plus ends usingfluorescence intensity measurements at the cortex. Error bars represent the SEM with a95% confidence interval, and n indicates the total number of measurements from 7-8embryos per condition. The t-test was used to determine statistical significance. ****P <0.0001.(C) Central confocal section of metaphase one-cell embryos expressing transgeneencoded mKate2::EBP-1, demonstrating the efficiency of ebp-1/3(RNAi). Images aremaximum intensity projections over time (10 images acquired every 300 ms). Note thatfor reasons that are not clear, mKate2::EBP-1 does not localize to MT plus ends. Scalebar, 5 um.(D) Schematic of the clip-1 locus. Mutations introduced by CRISPR-Cas9-based genomeediting to generate a null allele (clip-1) are indicated in black font.

Picture from Feng H et al. (2007) J Biol Chem "Properties, regulation, and in vivo functions of a novel protein kinase D: ...."

Figure 9. GATA enhancer sequences control dkf-2 gene promoter activity in intestinal cells in vivo. Transgenic C. elegans that express GFP-nuclear localization signal under control of the dkf-2 gene promoter were created (Experimental Procedures and Results). A presents a representative Nomarski interference image of an L2 larva. An arrowhead points to the tip of the head. Reporter (GFP) expression was monitored by confocal fluorescence microscopy (x600 magnification for embryos and x63 magnification for larvae). B, fluorescence signals reveal dkf-2 promoter activity in nuclei of intestinal cells. (Asterisks mark intestinal cell nuclei.) Note the outline of the sinuous intestinal lumen. dkf-2 promoter activity is evident in two cells near the posterior pharynx (arrow). C is a composite image generated by overlapping A and B. Composite images show dkf-2 promoter activity in the intestine of L1 (D) and embryonic (E) C. elegans. Reporter gene expression is also detected in pharyngeal cells in D and E (arrows). Deletion of GATA enhancer DNA sequences extinguished dkf-2 promoter activity in intestinal cells (e.g. in L2 larva (F), embryos (G), and adult animals (not shown)). GFP reporter expression in pharynx was not altered by loss of GATA enhancers (arrows, F and G). H shows the nucleotide sequence of proximal, 5' promoter/enhancer DNA and codons 1 and 2 of the dkf-2 gene. The initiator ATG codon is shown in italics and dashed underline; GATA enhancers are indicated with a solid underline; nucleotides deleted in the mutated promoter/enhancer DNA are marked with a bold font. The sequence is numbered according to the GenBank file (Z82052) for recombinant cosmid T25E12.

Picture from McDonald K et al. (2023) MicroPubl Biol "Using CRISPR knock-in of fluorescent tags to examine isoform-specific ...."

Figure 1. Comprehensive analysis of EGL-19 isoform expression:(A) The egl-19 locus is predicted to encode three isoforms of EGL-19 (a, b, and c). Exons are indicated as gray boxes linked by introns. The exon highlighted blue is unique to isoforms b and c. Black boxes represent the locations of the regions encoding the transmembrane domains that form the ion channel pore. Note that the truncated c isoform lacks these transmembrane domains. Fluorescent tags were inserted using CRISPR/Cas9 knock-in at the endogenous egl-19 locus. Colored boxes indicate the insertion location for each fluorescent tag relative to egl-19 gene structure (mSc = mScarlet and mNG = mNeonGreen). Orange box indicates the putative location of a neuron-specific promoter identified by the RegAtlas project (Serizay et al. 2020). Red font refers to RNAseq splice junction data that was obtained from WormBase (version WS288) and analyzed in this study. (B-G) Confocal imaging of EGL-19 tag expression. Yellow arrows indicate overlapping expression of mScarlet and GFP, revealing localization of the EGL-19(b) isoform. Red arrows indicate expression of GFP, revealing localization of the EGL-19(c) isoform. White arrows indicate punctate expression of mScarlet, which we hypothesize represents accumulation of mScarlet::EGL-19 in lysosomes. Pink arrows refer to a neuron which expresses both mNG and GFP, suggesting robust expression of EGL-19(c). Blue arrows refer to a bilateral pair of neurons which express mNG, but not GFP or mScarlet, suggesting more limited expression of EGL-19(c). (B) Representative images of co-expression of mScarlet::EGL-19(ab)/EGL-19(bc)::mNG in the pharyngeal bulb. Animals are F1 heterozygotes obtained from mating homozygous mScarlet::egl-19(ab) and egl-19(bc)::mNG parents. mNG expression in the pharyngeal muscle is so bright that it precludes analysis of mScarlet co-localization (compare mNG in B with GFP in E). (C-G) Representative images of co-expression of mScarlet::EGL-19(ab)/EGL-19(abc)::GFP in several different tissues. Animals are F1 heterozygotes obtained from mating homozygous mScarlet::egl-19(ab) and egl-19(abc)::GFP parents. mScarlet and GFP fluorescence levels are relatively balanced, allowing observation of overlapping expression in the "MERGE" column.