005f  (Expression Systems Inc)

Journal: bioRxiv

Article Title: Dredd-mediated cleavage of Kenny uncouples the IKK complex from selective autophagy to enable innate immunity

doi: 10.64898/2026.04.24.720600

Figure Lengend Snippet: (A) Drosophila S2 cells were transfected with empty vector, V5-tagged Dredd, and HA-tagged Kenny. The cell lysates were analysed by Western blotting using anti-HA, anti-V5, and anti-Actin antibodies, n>3. (B) Drosophila S2 cells were transfected with empty vector, HA-tagged Dredd and V5-tagged Kenny. V5-immunoprecipitations were performed and the samples were analysed by Western blotting with anti-HA, anti-V5, and anti-Actin antibodies, n=3. (C) Schematic representation of Dredd. (D) Drosophila S2 cells were transfected with empty vector, V5-tagged Kenny, and HA-tagged pro-domain or caspase domain of Dredd. V5-immunoprecipitations were performed and the samples were analysed by Western blotting with anti-HA, anti-V5, and anti-Actin antibodies, n=3. ( E ) Drosophila S2 cells were transfected with V5-tagged Kenny, and HA-tagged DED1 of Dredd. V5-immunoprecipitations were performed and the samples were analysed by Western blotting with anti-HA, anti-V5, and anti-Actin antibodies, n=3. (F) Schematic representation of the HA-tagged Kenny construct showing the positions of D21E, D27E, D67E and D88E point mutations. (G) Drosophila S2 cells were transfected with empty vector, V5-tagged Dredd, and HA-tagged Kenny WT and HA-tagged Kenny mutants D21E/D27E/D67E/D88E. The cell lysates were analysed by Western blotting using anti-HA, anti-V5, and anti-Actin antibodies, n>3.

Article Snippet: Drosophila Schneider S2 cells (Invitrogen) were grown at 25□°C using Schneider medium supplemented with 10% fetal bovine serum, 1% l-glutamine, and 0.5% penicillin/streptomycin.

Techniques: Transfection, Plasmid Preparation, Western Blot, Construct

Journal: bioRxiv

Article Title: Dredd-mediated cleavage of Kenny uncouples the IKK complex from selective autophagy to enable innate immunity

doi: 10.64898/2026.04.24.720600

Figure Lengend Snippet: (A) Structural modelling of the Dredd-Kenny interaction. Dredd (blue, AlphaFold: Q8IRY7) is modelled on the complex of two KSHV-FLIP (grey) proteins associated with a NEMO (pink) dimer (PDB: 3CL3). The molecular graphics and analyses were performed with the UCSF Chimera package . ( B ) Drosophila S2 cells were transfected with empty vector, HA-tagged wildtype, G98R, or C386A point mutant of Dredd and V5-tagged Kenny. V5-immunoprecipitations were performed and the samples were analysed by Western blotting with anti-HA, anti-V5, and anti-Actin antibodies. The ratio of immunoprecipitated Dredd to total Dredd was quantified, n=4. (C) Drosophila S2 cells were transfected with empty vector, V5-tagged wildtype, G98R, or C386A point mutant of Dredd and HA-tagged Kenny. Kenny cleavage was analysed from transfected S2 cells by Western blotting with anti-HA, anti-V5, and anti-Actin antibodies. The ratio of cleaved to total Kenny was quantified, n=5. (D) Adult male guts from Canton S flies or flies expressing GFP-Kenny ( NP1Gal4>UAS-GFP-Kenny ) in a wildtype, dredd D44 , or dredd L23 mutant background, fed with 5% sucrose and Ecc15 , were dissected. Kenny cleavage was analysed from dissected guts lysed in lysis buffer and analysed by Western blotting with anti-GFP and anti-Actin antibodies, n=3.

Article Snippet: Drosophila Schneider S2 cells (Invitrogen) were grown at 25□°C using Schneider medium supplemented with 10% fetal bovine serum, 1% l-glutamine, and 0.5% penicillin/streptomycin.

Techniques: Transfection, Plasmid Preparation, Mutagenesis, Western Blot, Immunoprecipitation, Expressing, Lysis

Journal: Life Science Alliance

Article Title: Flamenco plasticity tunes somatic piRNAs and rewires isoforms, with implications for heritable transposon spread

doi: 10.26508/lsa.202603650

Figure Lengend Snippet: (A) Schematic of the Springer element, showing the Gag, Pol, and Env ORFs and the structures of its unspliced and spliced transcripts. (B) Splice donor (red) and splice acceptor (blue) sites within Springer . (C) DNA sequence of the L3- Springer segment, extending from the 5′ end of the Env coding region to the 3′ end of L3- Springer . The Env TIS and stop codon are highlighted in red, and the L3- Springer –specific 29-nt deletion is indicated by a red triangle. The corresponding lowercase sequence is shown below. The PAS is shaded gray. (D) The L3- Springer element was introduced into S2 cells and ectopically expressed, and the resulting transcripts were examined for polyadenylation. All cDNA clones analyzed (n = 10) contained poly(A) tails.

Article Snippet: S2 cells were cultured at 26°C in Schneider’s Drosophila Medium (Gibco) with 10% FBS.

Techniques: Sequencing, Clone Assay

Journal: bioRxiv

Article Title: Togaram Ensures Axial Alignment of the Sperm Neck

doi: 10.64898/2026.04.15.718719

Figure Lengend Snippet: (A) Linear maps of human (top) and Drosophila (bottom) Togaram, Cep104, and CCDC66 depicting functional and structural domains. Domains are as indicated in the color key. (B) Both N– and C-terminally GFP-tagged Cep104 (green) binds Togaram (pink) and CCDC66 (blue). S2 cells were co-transfected with the indicated plasmids and anti-GFP co-IPs were prepared from cell lysates. Western blots of inputs and co-IPs were probed for GFP, Flag, and Myc. Arrow head colors correspond to indicated tagged protein colors. (C) Representative ExM images showing wild-type (left), cep104 null (center), and ccdc66 gRNA (right) Canoe stage spermatids with various alignment phenotypes. Spermatids were labeled for the nucleus (DAPI, blue; dotted line), the CA (Asl, magenta), and the centrioles (Ana1, green). Scale bar: ∼1 μm. (D) Quantification of wild-type (n=150), cep104 null (n=98), and ccdc66 gRNA (n=119) spermatids with various alignment phenotypes. (E) Working model in which Togaram, Cep104, and CCDC66 stabilize sperm head/neck microtubules to preserve a straight head-tail axis during the maintenance phase of spermiogenesis. Red question mark highlights a possible mechanism by which these proteins can directly regulate sperm neck stability independent of regulating microtubules.

Article Snippet: S2 cell expression vectors were generated by the Gateway cloning system (ThermoFisher Scientific) to create vectors with Actin5C promoter and the tags of interest (pAFHW, pAGW, pAMW, pAWG; F = Flag, G = GFP, M = Myc; https://emb.carnegiescience.edu/drosophila-gateway-vector-collection ). pAGW-MCS plasmid, expressing GFP alone was used as GFP control for IP experiments.

Techniques: Functional Assay, Transfection, Western Blot, Labeling