Journal: PLoS ONE

Article Title: Increased Apoptosis of Myoblasts in Drosophila Model for the Walker-Warburg Syndrome

doi: 10.1371/journal.pone.0011557

Figure Lengend Snippet: (A–D and I–L) wild type ( 1151 > GFPnls ). (E–H and M–P) tw mutant ( tw , 1151 > GFPnls ). (A) and (E) Images stained by anti-α-spectrin antibody. α-spectrin is a component of cytoskeleton inside of cell membrane and bind to actin. (B), (F), (J), and (N) Images stained by anti-DE-cadherin antibody. DE-cadherin is a cell adhesion molecule located in cell surface. (C), (G), (K), and (O) Nuclei of myoblasts. (D), (H), (L), and (P) Merged images of (A–C), (E–G), (I–K), and (M–O), respectively. (I) and (M) Images stained by anti-βPS-integrin antibody. βPS-integrin is a cell adhesion molecule located in cell surface and bind to extracellular matrix. TB, arrowheads, and arrows in (A), (E), (I), and (M) show tracheoblast, the region of myoblasts, and lateral region of myoblasts, respectively. The signals of α-spectrin and βPS-integrin excessively increased in the region of myoblasts compared to the lateral region of myoblasts in tw mutant although apoptosis increased in the myoblasts of tw mutant. These signals in wild type did not change between two regions. But we could not find any difference in the signal of DE-cadherin.

Article Snippet: The primary antibodies were used in the following dilutions: anti-cleaved caspase-3 (Asp175) rabbit polyclonal antibody, 1∶300 (Cell Signaling, http://www.cellsignal.com ); anti-phospho-histone H3 (Ser10) rabbit polyclonal antibody, 1∶100 (Millipore, http://www.millipore.com ); anti-α-spectrin mouse monoclonal antibody (3A9), 1∶25 (Developmental Studies Hybridoma Bank, http://dshb.biology.uiowa.edu/ ); anti-βPS-integrin mouse monoclonal antibody (CF.6G11), 1∶200 (Developmental Studies Hybridoma Bank); anti-DE-cadherin rat monoclonal antibody (DCAD2), 1∶25 (Developmental Studies Hybridoma Bank); and anti-Dg rabbit polyclonal antibody, 1∶100 .

Techniques: Mutagenesis, Staining, Membrane

Journal: bioRxiv

Article Title: Differential activation of JAK-STAT signaling in blood cell progenitors reveals functional compartmentalization of the Drosophila lymph gland

doi: 10.1101/2020.07.26.219717

Figure Lengend Snippet: (A, B) Primary lobes show increased expression of Pxn-GFP (green) in response to E . coli infection whereas PP are maintained. Graphs indicate ratio of Pxn + / DAPI + cells. Mann-Whitney nonparametric test was used for statistical analysis and error bars represent SEM. (C) Lymph gland response 2 days post-parasitism by L . boulardi . Immunostaining against the ß-integrin Myospheroid (yellow) reveal lamellocyte differentiation (red arrowhead). Phalloidin (white) marks actin. Green and orange inset: high magnification views of the primary and secondary lobes, respectively. Pri. indicates primary lobes, Sec. indicates secondary lobes and Tert. indicates tertiary lobes. Graphs indicate quantification for lamellocyte induction. * p<0 . 05 , ** p <0.01, *** p <0.001 and ns indicates non-significant. Scale bar: 100µm.

Article Snippet: Mouse anti-Antennapedia (1:20, DSHB #4C3), rat anti-DE-cadherin (1:10, DSHB #DCAD2), mouse anti- Ubx (1:20; DSHB #FP3.38), mouse anti-Dlp (1:50, DSHB #13G8), mouse anti-Myospheroid (1:50, DSHB #CF.6G11), mouse anti-P1 antibody (1:100, kind gift from Prof. Istvan Ando, Biological Research Center of the Hungarian Academy of Sciences), rabbit anti-GFP (1:1000, Clinisciences #TP401), chick or rabbit anti-GFP (1:500, Molecular Probes Inc.) rabbit anti-V5 (1:1000, ThermoFisher #PA1-993), mouse anti- ProPO antibody (1:5, Bioneeds), rabbit anti-Asrij (1:50, ( )).

Techniques: Expressing, Infection, MANN-WHITNEY, Immunostaining

Journal: PLoS ONE

Article Title: WNT5 Interacts with the Ryk Receptors Doughnut and Derailed to Mediate Muscle Attachment Site Selection in Drosophila melanogaster

doi: 10.1371/journal.pone.0032297

Figure Lengend Snippet: Third instar larval body walls of w 1118 ( A ), Wnt5 400 ( B ) and drl Red2 ( C ) mutant larvae are stained with anti-FAS2 (mAb 1D4). Wnt5 400 larvae and drl Red2 larvae frequently bypass their normal attachment sites and extend ventrally where they form new stable attachments. The original and ectopic tendons cells are indicated by + and *, respectively. FAS2 protein is evident at both sites. The penetrance of the bypass phenotypes is indicated in Table 1 . Anterior is up and ventral is left.

Article Snippet: The following primary antibodies were used on formaldehyde-fixed embryos or third instar larval body walls: anti-Muscle-Myosin mAb (Invitrogen), anti-FAS2 (1D4, Developmental Studies Hybridoma Bank (DSHB)), anti-βPS Integrin (CF.6G11; DSHB), guinea pig anti-SR (gift from T. Volk; ), rabbit-anti-GFP (Upstate), anti-Sex-Lethal ( ; DSHB), rabbit anti-MYC (Upstate) and affinity-purified rabbit anti-WNT5 .

Techniques: Mutagenesis, Staining

Journal: bioRxiv

Article Title: Relish plays a dynamic role in the niche to modulate Drosophila blood progenitor homeostasis in development and infection

doi: 10.1101/2021.04.04.438424

Figure Lengend Snippet: Genotypes of the larvae are mentioned in respective panels. Scale bar: 20µm A-A’ . Larval homogenates were spread on LB Agar plates to check the presence of commensal gut microbiota. In control scenario ( A ) bacterial colonies were visible post incubation whereas in axenic condition no growth was observed on the plates ( A’ ). B . The efficacy of removal of gut microflora was further checked by performing PCR analysis on DNA isolated from larval guts using 16S rDNA primers. Drosophila actin was used as control. Significant reduction in the amount of both Lactobacillus (compare lane 4 (axenic) with 1 (control) and Acetobacter (compare lane 5 (axenic) with 2 (control) species was observed in axenic condition compared to control scenario (compare lane 3 (axenic) and 6 (control). C-C’ . TRE-GFP expression in the hematopoietic niche (visualised by Antp, red) in axenic condition ( C’ ) is comparable to that of control ( C ). D-D’. Differentiation status (visualised by Hml>GFP , pan plasmatocyte marker) in axenic condition ( D’ ) is comparable to control ( D ). E-E’’ . Nuclear expression of Ecdysone receptor (red, EcR common) in the hematopoietic niche (green). F . Model depicting the developmental role of Relish in hematopoietic niche maintenance. Downregulation of Relish affects the proliferation and primary function of the niche by upregulated JNK signaling. Upregulated JNK disturbs niche homeostasis, thereby affecting progenitor maintenance. The white dotted line mark whole of the lymph gland in all cases. In all panels age of the larvae is 96 hrs AEH. The nuclei are marked with DAPI (Blue).

Article Snippet: Immunostaining and dissection (unless said otherwise) were performed using protocols described in ( ; ; ) using primary antibodies: mouse anti-c-Rel (1:50, a gift from N.Silverman , mouse anti Relish (1:50, 21F3, DSHB), mouse anti-Antp (1:10, 8C11, DSHB), mouse anti-Wg (1:3, 4D4, DSHB), mouse anti-P1 (1:40, a gift from I. Ando, rabbit anti-Ance (1:500, a gift from A. D. Shirras), rat anti-Ci (1:5, 2A1, DSHB), mouse anti-singed (1:20, Sn7C, DSHB), mouse anti-enabled (1:30, 5G2, DSHB), rabbit anti-PH3 (1:150, Cell signaling), rabbit anti-Hh (1:500, a gift from P. Ingham), mouse anti-Hindsight (1:5, 1G9, DSHB), mouse anti-EcR common (1:20, DDA2.7, DSHB), mouse anti-β-PS (1:3, CF.6G11, DSHB), rabbit-anti-GFP(1:100, 2555, Cell signalling), rat anti-shg (1:50, DCAD2, DSHB).

Techniques: Incubation, Isolation, Expressing, Marker