Journal: BMC biology

Article Title: Single-cell expression profile of Drosophila ovarian follicle stem cells illuminates spatial differentiation in the germarium.

doi: 10.1186/s12915-023-01636-9

Figure Lengend Snippet: Fig. 8 Expression patterns of selected genes. A–F Expression patterns of named gene products or derivatives in germaria co-stained with DAPI (blue in A–D) to reveal all nuclei and antibody to Fas3 (red in A–D; white in E–G) to allow location of the anterior edge of strong Fas3 expression (yellow arrows). A–A” Semaphorin 1a-GFP protein fusion (green) was detected on the surface of somatic cells; levels were low in region 1 (extreme anterior, left), peaked in region 2a ECs, FSCs, and early FCs, and declined in region 3 (posterior of the germarium) and the first budded egg chamber. B–B” Dystroglycan, detected by an antibody to the C-terminus (green), which connects the plasma membrane to ECM components, was seen lining the germarial wall from region 2a (white arrowheads in B’) and continuing posterior, consistent with its expected role and expression in all FSCs, early FCs, and a limited number of the most posterior ECs. Expression was also seen in the muscle sheath (cyan arrowheads in B). C–C” A Plum-GFP protein fusion (green) encoded by a large BAC genomic transgene was expressed most prominently in somatic cells, with highest levels in the FSC region and further posterior. D–D” A Cyclin B-GFP protein fusion showed occasional expression in layer1 FSCs (white arrow in D’ and D”) immediately anterior to the Fas3 border and was seen at similar levels in most FCs. Absence in some FCs is most likely due to protein degradation in G1 and S phases. Expression at the anterior of the germarium is within germline cells. E–G Santa Maria-Gal4 > UAS-RFP produced no RFP signal in most germaria. In other germaria, few cells expressed detectable RFP, including germaria with RFP signal in E a layer 1 FSC (white arrow) and a neighboring FC, F, G an FC adjacent to the Fas3 border, and G another early FC anterior to region 3. All bars, 20 µm

Article Snippet: Each ovary pair was then broken apart in PBS on a slide and mounted using DAPI fluoromountG mounting medium (Southern BioTech, OB010020).

Techniques: Expressing, Staining, Clinical Proteomics, Membrane, Produced

Journal: bioRxiv

Article Title: A divergent cyclic nucleotide binding protein promotes Plasmodium ookinete infection of the mosquito

doi: 10.1101/2025.01.30.635676

Figure Lengend Snippet: PbCBP-O is required for efficient midgut traversal. (A) PbΔCBP-O ookinetes show regular motility. Average speed is 3.07 and 3.2 μm/min for wild-type and ΔCBP-O, respectively (wild-type n = 38; PbΔCBP-O n = 48, unpaired t-test). (B) Comparable ookinete maturation in the mosquito midgut is observed in PbΔCBP-O and wildtype parasites. Nineteen or twenty-one hours post-infection, six midguts were scored per replicate (n = 2). Significance was analysed using an unpaired t-test. (C) Expression of micronemal proteins including CTRP, WARP and chitinase are unaffected in the absence of CBP-O. Actin is used as loading control (left panel). Chitinase secretion is maintained in PbΔCBP-O ookinetes. Western blot of culture supernatants from wild type and PbΔCBP-O ookinetes show similar levels of chitinase secretion (right panel). (D) Immunofluorescence showing decreased ookinete presence in the basal side of mosquito midguts. Midguts were fixed and stained with P25 (plasma membrane marker) and DAPI 24 hours post-infection. P25 + parasites were quantified, and single dots show number of ookinetes counted per midgut (n = 20 midguts per genotype, unpaired t-test.). Scale bar = 200μm. In all relevant panels, statistical significance is indicated as: ns = not significant and **** = p < 0.0001.

Article Snippet: The slides were mounted using 30μl of Fluoromount-G with DAPI (Fisher Scientific) and a 40mm coverslip and left overnight to set.

Techniques: Infection, Expressing, Control, Western Blot, Immunofluorescence, Staining, Membrane, Marker