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CRISPR/Cas9 tagging and expression of LtVtc1 and LtVtc4. (A) Schematic representation of LtVtc1 and LtVtc4. SPX: SYG1/Pho81/XPR domain, CD: Catalytic domain, TMH: Transmembrane helix domain. (B) PCR verification of mNG-tagged LtVtc1 and LtVtc4 showing a band at ∼2200 bp, absent in WT. (C) Fluorescent detection of mNG-LtLtVtc1, and LtVtc4 expressed in L. <t>tarentolae</t> ; Coomassie blue staining of WT L. tarentolae , mNG-LtVtc1, mNG-LtVtc4 confirms comparable total protein levels, ensuring consistent sample loading for fluorescent detection. (D) Fluorescence microscopy micrographs of mNG-LtLtVtc1 and mNG-LtVtc4 with mPPase in L.tarentolae (Pearson’s r = 0.85 and 0.83, respectively), scale bar: 5 µm.
L Tarentolae Parrot Tar, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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CRISPR/Cas9 tagging and expression of LtVtc1 and LtVtc4. (A) Schematic representation of LtVtc1 and LtVtc4. SPX: SYG1/Pho81/XPR domain, CD: Catalytic domain, TMH: Transmembrane helix domain. (B) PCR verification of mNG-tagged LtVtc1 and LtVtc4 showing a band at ∼2200 bp, absent in WT. (C) Fluorescent detection of mNG-LtLtVtc1, and LtVtc4 expressed in L. <t>tarentolae</t> ; Coomassie blue staining of WT L. tarentolae , mNG-LtVtc1, mNG-LtVtc4 confirms comparable total protein levels, ensuring consistent sample loading for fluorescent detection. (D) Fluorescence microscopy micrographs of mNG-LtLtVtc1 and mNG-LtVtc4 with mPPase in L.tarentolae (Pearson’s r = 0.85 and 0.83, respectively), scale bar: 5 µm.
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CRISPR/Cas9 tagging and expression of LtVtc1 and LtVtc4. (A) Schematic representation of LtVtc1 and LtVtc4. SPX: SYG1/Pho81/XPR domain, CD: Catalytic domain, TMH: Transmembrane helix domain. (B) PCR verification of mNG-tagged LtVtc1 and LtVtc4 showing a band at ∼2200 bp, absent in WT. (C) Fluorescent detection of mNG-LtLtVtc1, and LtVtc4 expressed in L. <t>tarentolae</t> ; Coomassie blue staining of WT L. tarentolae , mNG-LtVtc1, mNG-LtVtc4 confirms comparable total protein levels, ensuring consistent sample loading for fluorescent detection. (D) Fluorescence microscopy micrographs of mNG-LtLtVtc1 and mNG-LtVtc4 with mPPase in L.tarentolae (Pearson’s r = 0.85 and 0.83, respectively), scale bar: 5 µm.
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CRISPR/Cas9 tagging and expression of LtVtc1 and LtVtc4. (A) Schematic representation of LtVtc1 and LtVtc4. SPX: SYG1/Pho81/XPR domain, CD: Catalytic domain, TMH: Transmembrane helix domain. (B) PCR verification of mNG-tagged LtVtc1 and LtVtc4 showing a band at ∼2200 bp, absent in WT. (C) Fluorescent detection of mNG-LtLtVtc1, and LtVtc4 expressed in L. <t>tarentolae</t> ; Coomassie blue staining of WT L. tarentolae , mNG-LtVtc1, mNG-LtVtc4 confirms comparable total protein levels, ensuring consistent sample loading for fluorescent detection. (D) Fluorescence microscopy micrographs of mNG-LtLtVtc1 and mNG-LtVtc4 with mPPase in L.tarentolae (Pearson’s r = 0.85 and 0.83, respectively), scale bar: 5 µm.
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CRISPR/Cas9 tagging and expression of LtVtc1 and LtVtc4. (A) Schematic representation of LtVtc1 and LtVtc4. SPX: SYG1/Pho81/XPR domain, CD: Catalytic domain, TMH: Transmembrane helix domain. (B) PCR verification of mNG-tagged LtVtc1 and LtVtc4 showing a band at ∼2200 bp, absent in WT. (C) Fluorescent detection of mNG-LtLtVtc1, and LtVtc4 expressed in L. <t>tarentolae</t> ; Coomassie blue staining of WT L. tarentolae , mNG-LtVtc1, mNG-LtVtc4 confirms comparable total protein levels, ensuring consistent sample loading for fluorescent detection. (D) Fluorescence microscopy micrographs of mNG-LtLtVtc1 and mNG-LtVtc4 with mPPase in L.tarentolae (Pearson’s r = 0.85 and 0.83, respectively), scale bar: 5 µm.
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CRISPR/Cas9 tagging and expression of LtVtc1 and LtVtc4. (A) Schematic representation of LtVtc1 and LtVtc4. SPX: SYG1/Pho81/XPR domain, CD: Catalytic domain, TMH: Transmembrane helix domain. (B) PCR verification of mNG-tagged LtVtc1 and LtVtc4 showing a band at ∼2200 bp, absent in WT. (C) Fluorescent detection of mNG-LtLtVtc1, and LtVtc4 expressed in L. <t>tarentolae</t> ; Coomassie blue staining of WT L. tarentolae , mNG-LtVtc1, mNG-LtVtc4 confirms comparable total protein levels, ensuring consistent sample loading for fluorescent detection. (D) Fluorescence microscopy micrographs of mNG-LtLtVtc1 and mNG-LtVtc4 with mPPase in L.tarentolae (Pearson’s r = 0.85 and 0.83, respectively), scale bar: 5 µm.
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CRISPR/Cas9 tagging and expression of LtVtc1 and LtVtc4. (A) Schematic representation of LtVtc1 and LtVtc4. SPX: SYG1/Pho81/XPR domain, CD: Catalytic domain, TMH: Transmembrane helix domain. (B) PCR verification of mNG-tagged LtVtc1 and LtVtc4 showing a band at ∼2200 bp, absent in WT. (C) Fluorescent detection of mNG-LtLtVtc1, and LtVtc4 expressed in L. <t>tarentolae</t> ; Coomassie blue staining of WT L. tarentolae , mNG-LtVtc1, mNG-LtVtc4 confirms comparable total protein levels, ensuring consistent sample loading for fluorescent detection. (D) Fluorescence microscopy micrographs of mNG-LtLtVtc1 and mNG-LtVtc4 with mPPase in L.tarentolae (Pearson’s r = 0.85 and 0.83, respectively), scale bar: 5 µm.
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CRISPR/Cas9 tagging and expression of LtVtc1 and LtVtc4. (A) Schematic representation of LtVtc1 and LtVtc4. SPX: SYG1/Pho81/XPR domain, CD: Catalytic domain, TMH: Transmembrane helix domain. (B) PCR verification of mNG-tagged LtVtc1 and LtVtc4 showing a band at ∼2200 bp, absent in WT. (C) Fluorescent detection of mNG-LtLtVtc1, and LtVtc4 expressed in L. <t>tarentolae</t> ; Coomassie blue staining of WT L. tarentolae , mNG-LtVtc1, mNG-LtVtc4 confirms comparable total protein levels, ensuring consistent sample loading for fluorescent detection. (D) Fluorescence microscopy micrographs of mNG-LtLtVtc1 and mNG-LtVtc4 with mPPase in L.tarentolae (Pearson’s r = 0.85 and 0.83, respectively), scale bar: 5 µm.
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Cosmo Bio USA primary antibodies against phosphorylated tar dna- binding protein 43 kda (ptdp43)
CRISPR/Cas9 tagging and expression of LtVtc1 and LtVtc4. (A) Schematic representation of LtVtc1 and LtVtc4. SPX: SYG1/Pho81/XPR domain, CD: Catalytic domain, TMH: Transmembrane helix domain. (B) PCR verification of mNG-tagged LtVtc1 and LtVtc4 showing a band at ∼2200 bp, absent in WT. (C) Fluorescent detection of mNG-LtLtVtc1, and LtVtc4 expressed in L. <t>tarentolae</t> ; Coomassie blue staining of WT L. tarentolae , mNG-LtVtc1, mNG-LtVtc4 confirms comparable total protein levels, ensuring consistent sample loading for fluorescent detection. (D) Fluorescence microscopy micrographs of mNG-LtLtVtc1 and mNG-LtVtc4 with mPPase in L.tarentolae (Pearson’s r = 0.85 and 0.83, respectively), scale bar: 5 µm.
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CRISPR/Cas9 tagging and expression of LtVtc1 and LtVtc4. (A) Schematic representation of LtVtc1 and LtVtc4. SPX: SYG1/Pho81/XPR domain, CD: Catalytic domain, TMH: Transmembrane helix domain. (B) PCR verification of mNG-tagged LtVtc1 and LtVtc4 showing a band at ∼2200 bp, absent in WT. (C) Fluorescent detection of mNG-LtLtVtc1, and LtVtc4 expressed in L. <t>tarentolae</t> ; Coomassie blue staining of WT L. tarentolae , mNG-LtVtc1, mNG-LtVtc4 confirms comparable total protein levels, ensuring consistent sample loading for fluorescent detection. (D) Fluorescence microscopy micrographs of mNG-LtLtVtc1 and mNG-LtVtc4 with mPPase in L.tarentolae (Pearson’s r = 0.85 and 0.83, respectively), scale bar: 5 µm.
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CRISPR/Cas9 tagging and expression of LtVtc1 and LtVtc4. (A) Schematic representation of LtVtc1 and LtVtc4. SPX: SYG1/Pho81/XPR domain, CD: Catalytic domain, TMH: Transmembrane helix domain. (B) PCR verification of mNG-tagged LtVtc1 and LtVtc4 showing a band at ∼2200 bp, absent in WT. (C) Fluorescent detection of mNG-LtLtVtc1, and LtVtc4 expressed in L. tarentolae ; Coomassie blue staining of WT L. tarentolae , mNG-LtVtc1, mNG-LtVtc4 confirms comparable total protein levels, ensuring consistent sample loading for fluorescent detection. (D) Fluorescence microscopy micrographs of mNG-LtLtVtc1 and mNG-LtVtc4 with mPPase in L.tarentolae (Pearson’s r = 0.85 and 0.83, respectively), scale bar: 5 µm.

Journal: bioRxiv

Article Title: Novel Binding Partners of the Vacuolar Transporter Chaperone (VTC) complex in Acidocalcisomes of Leishmania tarentolae

doi: 10.1101/2025.09.23.677757

Figure Lengend Snippet: CRISPR/Cas9 tagging and expression of LtVtc1 and LtVtc4. (A) Schematic representation of LtVtc1 and LtVtc4. SPX: SYG1/Pho81/XPR domain, CD: Catalytic domain, TMH: Transmembrane helix domain. (B) PCR verification of mNG-tagged LtVtc1 and LtVtc4 showing a band at ∼2200 bp, absent in WT. (C) Fluorescent detection of mNG-LtLtVtc1, and LtVtc4 expressed in L. tarentolae ; Coomassie blue staining of WT L. tarentolae , mNG-LtVtc1, mNG-LtVtc4 confirms comparable total protein levels, ensuring consistent sample loading for fluorescent detection. (D) Fluorescence microscopy micrographs of mNG-LtLtVtc1 and mNG-LtVtc4 with mPPase in L.tarentolae (Pearson’s r = 0.85 and 0.83, respectively), scale bar: 5 µm.

Article Snippet: GO analysis of biological processes, cell components, and molecular functions was performed on the 412 overlapping LtVtc1 and LtVtc4 interactors using the TriTrypDB GO analysis tool [ ] with the complete L. tarentolae parrot tar II 2019 (ATCC 30267) genome as the reference.

Techniques: CRISPR, Expressing, Staining, Fluorescence, Microscopy

Colocalisation analysis of L. tarentolae expressing mNG-LtVtc4 and mCh-tagged by immunofluorescence microscopy. mNG-LtVtc4 colocalises with mCh-LtVtc1 (Pearson’s r = 0.83) as well as with mCh-LtPho91, mCh-LtCa²⁺-ATPase, mCh-LtV-H⁺-ATPase_A, and mCh-LtZnT, showing Pearson’s correlation coefficients of 0.87, 0.83, 0.87, and 0.84, respectively. A lower degree of colocalisation is observed with mCh-IP₃R (r = 0.67). Scale bar: 5 µm.

Journal: bioRxiv

Article Title: Novel Binding Partners of the Vacuolar Transporter Chaperone (VTC) complex in Acidocalcisomes of Leishmania tarentolae

doi: 10.1101/2025.09.23.677757

Figure Lengend Snippet: Colocalisation analysis of L. tarentolae expressing mNG-LtVtc4 and mCh-tagged by immunofluorescence microscopy. mNG-LtVtc4 colocalises with mCh-LtVtc1 (Pearson’s r = 0.83) as well as with mCh-LtPho91, mCh-LtCa²⁺-ATPase, mCh-LtV-H⁺-ATPase_A, and mCh-LtZnT, showing Pearson’s correlation coefficients of 0.87, 0.83, 0.87, and 0.84, respectively. A lower degree of colocalisation is observed with mCh-IP₃R (r = 0.67). Scale bar: 5 µm.

Article Snippet: GO analysis of biological processes, cell components, and molecular functions was performed on the 412 overlapping LtVtc1 and LtVtc4 interactors using the TriTrypDB GO analysis tool [ ] with the complete L. tarentolae parrot tar II 2019 (ATCC 30267) genome as the reference.

Techniques: Expressing, Immunofluorescence, Microscopy

Journal: bioRxiv

Article Title: Novel Binding Partners of the Vacuolar Transporter Chaperone (VTC) complex in Acidocalcisomes of Leishmania tarentolae

doi: 10.1101/2025.09.23.677757

Figure Lengend Snippet:

Article Snippet: GO analysis of biological processes, cell components, and molecular functions was performed on the 412 overlapping LtVtc1 and LtVtc4 interactors using the TriTrypDB GO analysis tool [ ] with the complete L. tarentolae parrot tar II 2019 (ATCC 30267) genome as the reference.

Techniques: Binding Assay

Colocalisation analysis of L. tarentolae expressing mNG-LtVtc4 and mCh-tagged LtVBP1, LtVBP2, and LtVBP3 by immunofluorescence microscopy. All three proteins show colocalisation with LtVtc4 in acidocalcisomes. Pearson’s correlation coefficients with LtVtc4 are 0.83 for LtVBP1, 0.85 for LtVBP2, and 0.82 for LtVBP3. Scale bar: 5 µm.

Journal: bioRxiv

Article Title: Novel Binding Partners of the Vacuolar Transporter Chaperone (VTC) complex in Acidocalcisomes of Leishmania tarentolae

doi: 10.1101/2025.09.23.677757

Figure Lengend Snippet: Colocalisation analysis of L. tarentolae expressing mNG-LtVtc4 and mCh-tagged LtVBP1, LtVBP2, and LtVBP3 by immunofluorescence microscopy. All three proteins show colocalisation with LtVtc4 in acidocalcisomes. Pearson’s correlation coefficients with LtVtc4 are 0.83 for LtVBP1, 0.85 for LtVBP2, and 0.82 for LtVBP3. Scale bar: 5 µm.

Article Snippet: GO analysis of biological processes, cell components, and molecular functions was performed on the 412 overlapping LtVtc1 and LtVtc4 interactors using the TriTrypDB GO analysis tool [ ] with the complete L. tarentolae parrot tar II 2019 (ATCC 30267) genome as the reference.

Techniques: Expressing, Immunofluorescence, Microscopy

Pulldown assay of novel VTC binding partners with mNG-LtVtc4 in L. tarentolae. (A-C) SDS-PAGE with fluorescence detection shows capture of mCh-LtVBP1, mCh-LtVBP2, and mCh-LtVBP3 by mNG-LtVtc4 using mNeonGreen-Trap agarose beads, as indicated by signal in the bound fraction. (D) Pulldown of mNG-LtVtc4 with mCh-LtVBP3 using ChromoTek RFP-Trap beads confirms interaction. Lane labels: I – input; FT – flow-through; W3 – third wash; B – bound fraction. Green bands correspond to mNG-LtVtc4; some I and B lines appear as yellowish due to partial detection of the mNG tag in the 532 nm channel ( S6 Fig ). Red bands correspond to mCherry-tagged proteins. The fluorescence signal marked by a white asterisk in the gel represents free mCh tag, visible only in the input due to partial degradation.

Journal: bioRxiv

Article Title: Novel Binding Partners of the Vacuolar Transporter Chaperone (VTC) complex in Acidocalcisomes of Leishmania tarentolae

doi: 10.1101/2025.09.23.677757

Figure Lengend Snippet: Pulldown assay of novel VTC binding partners with mNG-LtVtc4 in L. tarentolae. (A-C) SDS-PAGE with fluorescence detection shows capture of mCh-LtVBP1, mCh-LtVBP2, and mCh-LtVBP3 by mNG-LtVtc4 using mNeonGreen-Trap agarose beads, as indicated by signal in the bound fraction. (D) Pulldown of mNG-LtVtc4 with mCh-LtVBP3 using ChromoTek RFP-Trap beads confirms interaction. Lane labels: I – input; FT – flow-through; W3 – third wash; B – bound fraction. Green bands correspond to mNG-LtVtc4; some I and B lines appear as yellowish due to partial detection of the mNG tag in the 532 nm channel ( S6 Fig ). Red bands correspond to mCherry-tagged proteins. The fluorescence signal marked by a white asterisk in the gel represents free mCh tag, visible only in the input due to partial degradation.

Article Snippet: GO analysis of biological processes, cell components, and molecular functions was performed on the 412 overlapping LtVtc1 and LtVtc4 interactors using the TriTrypDB GO analysis tool [ ] with the complete L. tarentolae parrot tar II 2019 (ATCC 30267) genome as the reference.

Techniques: Binding Assay, SDS Page, Fluorescence