A novel lineage-tracing mouse model for studying early MmuPV1 infections
- Department of Biological Sciences, University of Cyprus, Cyprus
- Department of Cancer Genetics, Therapeutics & Ultrastructural Pathology, The Cyprus Institute of Neurology & Genetics, Cyprus
Figures
Figure 1
Cre expression from MmuPV1-lox-Cre-lox leads to plasmid recombination in vitro.
(A) Immunofluorescence of mouse embryonic fibroblasts (MEFs) (isolated from R26R-EYFP mice) transfected with pBABE-GFP or MmuPV1-lox-Cre-lox (MmuPV1-Cre) plasmids 48 hr post-transfection (green: …
Figure 2
Cells which taken up MmuPV1-lox-Cre-lox plasmid can be detected and traced over time in vivo.
(A) Experimental design; the MmuPV1-lox-Cre-lox plasmid is delivered to the tail skin of R26R-lox-STOP-lox-eYFP mice (of pure C57BL/6 genetic background) after a UV-B irradiation and a superficial …
Figure 3 with 1 supplement
Tissues with MmuPV1-lox-Cre-lox plasmid express viral transcripts in vivo.
(A–B) RNAscope of the MmuPV1-lox-Cre-lox plasmid delivered tail sections of the R26R-lox-STOP-lox-eYFP mice at the indicated time points to assess the presence of viral transcripts E6/E7 and E1/E4, …
Figure 3—figure supplement 1
RNAscope controls with DNAse and/or RNAse and CAG-Cre controls for the detection of MmuPV1-L1 and Cre loss sequences.
(A–B) RNAscope of the MmuPV1-Cre plasmid delivered tail sections of the Rosa26-lox-stop-lox-YFP mice at 5 days post-delivery to assess the presence of viral transcripts E6/E7 (A) and E1/E4 (B), …
Figure 4 with 2 supplements
Cells with MmuPV1-lox-Cre-lox plasmid have increased proliferation rate and decreased MHC-I expression on the cell surface.
(A) Percentages of YFP+ cells. (B) Percentages of Ki-67+ cells out of only YFP+ cells. (C) Percentages of MHC-I+ cells out of only YFP+ cells. Mock infection with PBS (red square) was only performed …
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Figure 4—source data 1
The source data of FACS for Figure 4 and its supplements is included in a Microsoft Excel spreadsheet called “Source data - FACS”.
- https://cdn.elifesciences.org/articles/72638/elife-72638-fig4-data1-v1.xlsx
Figure 4—figure supplement 1
Cells with either MmuPV1-Cre or CAG-Cre control plasmid can be detected and analyzed by flow cytometry.
(A) The population of cells was detected depending on their size and complexity, omitting the dead cells (forward scatter (FSC)-side scatter (SSC) gate) and later excluding the doublets …
Figure 4—figure supplement 2
YFP- cells in the tissues treated with MmuPV1-Cre and CAG-Cre control plasmid have similar Ki-67 and MHC-I expression.
(A) Absolute numbers (left panel) and percentages (right panel) of Ki-67+ cells out of only YFP- cells. (B) Absolute numbers (left panel) and percentages (right panel) of MHC-I+ cells out of only …
Additional files
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Source data 1
Source data for gels in Figures 1 and 3, Figure 3—figure supplement 1 are provided in a ZIP file called “Source data Gels”.
Source data for sequences and plasmid creation in detail are provided in a ZIP file called “Source data Sequences and Plasmid creation”. The source data of FACS for Figure 4 and its supplements is included in a Microsoft Excel spreadsheet called “Source data - FACS”.
- https://cdn.elifesciences.org/articles/72638/elife-72638-data1-v1.zip
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Source data 2
Source data for sequences and plasmid creation in detail are provided in a ZIP file called “Source data Sequences and Plasmid creation”.
- https://cdn.elifesciences.org/articles/72638/elife-72638-data2-v1.zip
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Transparent reporting form
- https://cdn.elifesciences.org/articles/72638/elife-72638-transrepform1-v1.pdf
