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(A) Diagram of Glycine2 and Serines in Wek tested. (B) Mass spectrometry of purified Wek-FLAG: S2 cells were co-transfected with wek-FLAG and Toll-6HA, or wek-FLAG and pelle-HA and stimulated with purified DNT-2, Wek was purified, and found to be phosphorylated at Ser164. This Ser was not phosphorylated in controls. (C,D) Phostag gel of wek mutant forms expressed in S2 cells, showing: (C) western blot input; (D) Phostag gel, showing the distinct migration profile of the Wek mutant forms. Asterisks indicate bands modified by the co-transfection with Pelle. (E,F) <t>Microscopy</t> from transfected S2 cells showing that site directed mutagenesis of wek in G2A, S164A and S168A, alter the subcellular distribution of Wek, and this is further influenced by pelle . Wek visualised with anti-FLAG antibodies (green) and nuclei with DAPI (magenta). (G) Illustration showing that phosphorylation at Ser6 and Ser168 are required for the nuclear translocation of Wek. Scale bar: (E,F) 10μm
Laser Scanning Confocal Microscopy Image Data, supplied by Oxford Instruments, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A) Diagram of Glycine2 and Serines in Wek tested. (B) Mass spectrometry of purified Wek-FLAG: S2 cells were co-transfected with wek-FLAG and Toll-6HA, or wek-FLAG and pelle-HA and stimulated with purified DNT-2, Wek was purified, and found to be phosphorylated at Ser164. This Ser was not phosphorylated in controls. (C,D) Phostag gel of wek mutant forms expressed in S2 cells, showing: (C) western blot input; (D) Phostag gel, showing the distinct migration profile of the Wek mutant forms. Asterisks indicate bands modified by the co-transfection with Pelle. (E,F) <t>Microscopy</t> from transfected S2 cells showing that site directed mutagenesis of wek in G2A, S164A and S168A, alter the subcellular distribution of Wek, and this is further influenced by pelle . Wek visualised with anti-FLAG antibodies (green) and nuclei with DAPI (magenta). (G) Illustration showing that phosphorylation at Ser6 and Ser168 are required for the nuclear translocation of Wek. Scale bar: (E,F) 10μm
High Speed Confocal Microscopy, supplied by Oxford Instruments, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A) Diagram of Glycine2 and Serines in Wek tested. (B) Mass spectrometry of purified Wek-FLAG: S2 cells were co-transfected with wek-FLAG and Toll-6HA, or wek-FLAG and pelle-HA and stimulated with purified DNT-2, Wek was purified, and found to be phosphorylated at Ser164. This Ser was not phosphorylated in controls. (C,D) Phostag gel of wek mutant forms expressed in S2 cells, showing: (C) western blot input; (D) Phostag gel, showing the distinct migration profile of the Wek mutant forms. Asterisks indicate bands modified by the co-transfection with Pelle. (E,F) <t>Microscopy</t> from transfected S2 cells showing that site directed mutagenesis of wek in G2A, S164A and S168A, alter the subcellular distribution of Wek, and this is further influenced by pelle . Wek visualised with anti-FLAG antibodies (green) and nuclei with DAPI (magenta). (G) Illustration showing that phosphorylation at Ser6 and Ser168 are required for the nuclear translocation of Wek. Scale bar: (E,F) 10μm
Spinning Disk Confocal Microscopy, supplied by Yokogawa Electric, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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spinning disk confocal microscopy  (Oxford Instruments)
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(A) Diagram of Glycine2 and Serines in Wek tested. (B) Mass spectrometry of purified Wek-FLAG: S2 cells were co-transfected with wek-FLAG and Toll-6HA, or wek-FLAG and pelle-HA and stimulated with purified DNT-2, Wek was purified, and found to be phosphorylated at Ser164. This Ser was not phosphorylated in controls. (C,D) Phostag gel of wek mutant forms expressed in S2 cells, showing: (C) western blot input; (D) Phostag gel, showing the distinct migration profile of the Wek mutant forms. Asterisks indicate bands modified by the co-transfection with Pelle. (E,F) <t>Microscopy</t> from transfected S2 cells showing that site directed mutagenesis of wek in G2A, S164A and S168A, alter the subcellular distribution of Wek, and this is further influenced by pelle . Wek visualised with anti-FLAG antibodies (green) and nuclei with DAPI (magenta). (G) Illustration showing that phosphorylation at Ser6 and Ser168 are required for the nuclear translocation of Wek. Scale bar: (E,F) 10μm
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laser scanning confocal microscopy  (Oxford Instruments)
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(A) Diagram of Glycine2 and Serines in Wek tested. (B) Mass spectrometry of purified Wek-FLAG: S2 cells were co-transfected with wek-FLAG and Toll-6HA, or wek-FLAG and pelle-HA and stimulated with purified DNT-2, Wek was purified, and found to be phosphorylated at Ser164. This Ser was not phosphorylated in controls. (C,D) Phostag gel of wek mutant forms expressed in S2 cells, showing: (C) western blot input; (D) Phostag gel, showing the distinct migration profile of the Wek mutant forms. Asterisks indicate bands modified by the co-transfection with Pelle. (E,F) <t>Microscopy</t> from transfected S2 cells showing that site directed mutagenesis of wek in G2A, S164A and S168A, alter the subcellular distribution of Wek, and this is further influenced by pelle . Wek visualised with anti-FLAG antibodies (green) and nuclei with DAPI (magenta). (G) Illustration showing that phosphorylation at Ser6 and Ser168 are required for the nuclear translocation of Wek. Scale bar: (E,F) 10μm
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(A) Diagram of Glycine2 and Serines in Wek tested. (B) Mass spectrometry of purified Wek-FLAG: S2 cells were co-transfected with wek-FLAG and Toll-6HA, or wek-FLAG and pelle-HA and stimulated with purified DNT-2, Wek was purified, and found to be phosphorylated at Ser164. This Ser was not phosphorylated in controls. (C,D) Phostag gel of wek mutant forms expressed in S2 cells, showing: (C) western blot input; (D) Phostag gel, showing the distinct migration profile of the Wek mutant forms. Asterisks indicate bands modified by the co-transfection with Pelle. (E,F) <t>Microscopy</t> from transfected S2 cells showing that site directed mutagenesis of wek in G2A, S164A and S168A, alter the subcellular distribution of Wek, and this is further influenced by pelle . Wek visualised with anti-FLAG antibodies (green) and nuclei with DAPI (magenta). (G) Illustration showing that phosphorylation at Ser6 and Ser168 are required for the nuclear translocation of Wek. Scale bar: (E,F) 10μm
Bc43 Confocal Microscopy Imaging System, supplied by Oxford Instruments, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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high resolution confocal microscopy  (Oxford Instruments)
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Oxford Instruments high resolution confocal microscopy
(A) Diagram of Glycine2 and Serines in Wek tested. (B) Mass spectrometry of purified Wek-FLAG: S2 cells were co-transfected with wek-FLAG and Toll-6HA, or wek-FLAG and pelle-HA and stimulated with purified DNT-2, Wek was purified, and found to be phosphorylated at Ser164. This Ser was not phosphorylated in controls. (C,D) Phostag gel of wek mutant forms expressed in S2 cells, showing: (C) western blot input; (D) Phostag gel, showing the distinct migration profile of the Wek mutant forms. Asterisks indicate bands modified by the co-transfection with Pelle. (E,F) <t>Microscopy</t> from transfected S2 cells showing that site directed mutagenesis of wek in G2A, S164A and S168A, alter the subcellular distribution of Wek, and this is further influenced by pelle . Wek visualised with anti-FLAG antibodies (green) and nuclei with DAPI (magenta). (G) Illustration showing that phosphorylation at Ser6 and Ser168 are required for the nuclear translocation of Wek. Scale bar: (E,F) 10μm
High Resolution Confocal Microscopy, supplied by Oxford Instruments, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A) Diagram of Glycine2 and Serines in Wek tested. (B) Mass spectrometry of purified Wek-FLAG: S2 cells were co-transfected with wek-FLAG and Toll-6HA, or wek-FLAG and pelle-HA and stimulated with purified DNT-2, Wek was purified, and found to be phosphorylated at Ser164. This Ser was not phosphorylated in controls. (C,D) Phostag gel of wek mutant forms expressed in S2 cells, showing: (C) western blot input; (D) Phostag gel, showing the distinct migration profile of the Wek mutant forms. Asterisks indicate bands modified by the co-transfection with Pelle. (E,F) Microscopy from transfected S2 cells showing that site directed mutagenesis of wek in G2A, S164A and S168A, alter the subcellular distribution of Wek, and this is further influenced by pelle . Wek visualised with anti-FLAG antibodies (green) and nuclei with DAPI (magenta). (G) Illustration showing that phosphorylation at Ser6 and Ser168 are required for the nuclear translocation of Wek. Scale bar: (E,F) 10μm

Journal: bioRxiv

Article Title: Weckle is a molecular switch that diverts Toll signalling from innate immunity towards growth by engaging Yki

doi: 10.64898/2026.02.18.706625

Figure Lengend Snippet: (A) Diagram of Glycine2 and Serines in Wek tested. (B) Mass spectrometry of purified Wek-FLAG: S2 cells were co-transfected with wek-FLAG and Toll-6HA, or wek-FLAG and pelle-HA and stimulated with purified DNT-2, Wek was purified, and found to be phosphorylated at Ser164. This Ser was not phosphorylated in controls. (C,D) Phostag gel of wek mutant forms expressed in S2 cells, showing: (C) western blot input; (D) Phostag gel, showing the distinct migration profile of the Wek mutant forms. Asterisks indicate bands modified by the co-transfection with Pelle. (E,F) Microscopy from transfected S2 cells showing that site directed mutagenesis of wek in G2A, S164A and S168A, alter the subcellular distribution of Wek, and this is further influenced by pelle . Wek visualised with anti-FLAG antibodies (green) and nuclei with DAPI (magenta). (G) Illustration showing that phosphorylation at Ser6 and Ser168 are required for the nuclear translocation of Wek. Scale bar: (E,F) 10μm

Article Snippet: To analyse glial cell membrane volume in adult antennal lobes, laser scanning confocal microscopy image data were processed with Imaris using the “Surface” module.

Techniques: Mass Spectrometry, Purification, Transfection, Mutagenesis, Western Blot, Migration, Modification, Cotransfection, Microscopy, Phospho-proteomics, Translocation Assay