Additional file 1: of “Salivary gland cellular architecture in the Asian malaria vector mosquito Anopheles stephensi”
Creators: name: Wells, Michael
name: Andrew, Deborah
Publisher: name: Figshare
Year: 2015
Subjects: subject: Biochemistry
subject: Microbiology
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: Cell Biology
subject: Genetics
subject: Molecular Biology
subject: Physiology
subject: Evolutionary Biology
subject: Immunology
subject: FOS: Clinical medicine
subject: FOS: Clinical medicine
subject: Developmental Biology
subject: 110309 Infectious Diseases
subject: FOS: Health sciences
subject: FOS: Health sciences
subject: Plant Biology

description: Figure S1. Adult Anopheles stephensi salivary gland lobe and cellular morphometry. (A) Graph of average length (blue) and width (red) of each lobe from An. stephensi. Ten samples were measured per lobe. Results indicate that lobe length varies more than lobe width, but variations are similar across lobes (standard deviations are shown on the graph). (B) Graph of adult An. stephensi SG cellular morphometry. Each data point represents 50 cellular measurements among 10 lobes. Results show low variability in both female and male gland cellular dimensions, except for proximal lateral cell length, which varies with asymmetric positioning of the duct within that part of the gland. (C) Cross-sections from Nile Red and DAPI stained SGs illustrating the number of nuclei in circumference in the female distal lateral (DL), proximal lateral (PL), and medial lobe (M), and male gland. Proximal (P) and distal (D) are labeled on each lobe/gland for orientation. Arrow in (PL) marks the PL cross-section. The number of circumferential nuclei varies along the length of each lobe. These cell shape and duct observations agree well with previously published electron microscopy cross-sectional views [11]. Scale bars are 50 microns long. Figure S2. Additional details regarding adult Drosophila salivary gland cellular architecture. Adult D. melanogaster (Canton S) SGs stained with DAPI (nuclei), phalloidin (actin), and WGA (chitin; A) or Nile Red (lipids; B-D). A) Evidence of cell turnover was observed, including DNA fragmentation (white arrow), basal cell detachment, and actin accumulation (yellow arrow). (B-D) Female (B) and Male (C-D) SG cell shape. Results show Nile Red staining throughout the cell membranes, highlighting the cuboidal shape of the secretory cells (e.g. Bii, white arrows). Actin is well organized into apico-basal fibers (Biii), and debris can sometimes be visualized in the lumen (Biv, white arrow). The lumenal surface appears irregular [compare Cii to D (proximal portion from another gland)]. Images not labeled with “MIP” (maximum intensity projections), are single slices. D is a MIP of two slices. Scale bar lengths are: 50 microns—A, Bi, Ci; 20 microns—Bii, Biii, Biv, Cii, D. Figure S3. Larval Drosophila salivary gland nuclear dimension correlate with known ploidy. (A) Male D. melanogaster (Oregon R) larval SGs stained with DAPI (nuclei). Three classes of nuclei are visible: 1) imaginal nuclei (blue arrow), 2) duct nuclei (red arrow), and 3) secretory nuclei (green arrow). Imaginal nuclei are known to be diploid; duct nuclei are weakly polyploid and secretory nuclei are very highly polyploid (polytene). (B) Graph of nuclear diameters of these three cell types. Sample size is given at the base of each bar. Note that nuclear size increases with an increase in ploidy. (C) An. stephensi nuclear diameter measurements. A graph (Ci) of nuclear diameter measurements from two populations of cells (duct, and secretory cells), reveals that both cell types had consistently sized nuclei; secretory cell nuclei are a little more than twice the diameter of duct nuclei. Secretory cell and duct cell measurements were taken from Fig. 2a and similar images. Small DAPI body (micronucleus) measurements were taken from Cii. In this female DL lobe, micronuclei are abundant (sites of enriched DAPI and Nile Red colocalization). Micronucleus morphology was often elongated, not spherical, so maximal length was measured. Micronucleus length distribution is plotted in Ciii, sorted from smallest to largest. Discontinuous outlier values are boxed. (D) Visual comparison of D. melanogaster imaginal cell (Di) and An. stephensi micronucleus (Dii) size. Images are the same scale, and nuclei of interest are marked by white arrows. *: p-value