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    • carcinoma derived cell lines siha  (ATCC)
    98
    ATCC carcinoma derived cell lines siha
    Expression profiles <t>of</t> <t>HPV16</t> E6 and E6*I in stable cell lines. ( A ) Western blotting analysis showing p53, E6 and β-actin protein levels in U-2 OS parental cell line (U-CTL#par) and in stably transfected cells with pXJ40 empty vector (U-CTL #1 and #3) or pXJ40-E6All vector (U-E6All #1, #6 and #7) or pXJ40-E6*I vector (U-E6*I #1, #6 and #14). β-actin is used as loading control. Full-length blots are presented in Figure . ( B ) Western blotting analysis comparing protein levels of E6 and p53 between U-2 OS cellular clones (U-CTL#par, U-E6All #1, #6 and #7) and naturally HPV16-infected cells (W12_20861, W12_20862, W12_20863, <t>SiHa</t> and Ca Ski cell lines). β-actin is used as loading control. (C) RT-PCR analysis of alternatively spliced HPV16 E6 transcripts in U-E6All and U-E6*I stably transfected cells. As expected no E6 transcripts were observed in U-2 OS parental cell line (U-CTL#par) and in U-CTL #1 and #3 stably transfected cells. The detection of the vector in stably transfected cells was done by pXJ40 vector specific primers and RPLP0 was used as loading control. Full-length gels are presented in Figure . (D) RT-PCR analysis of alternatively spliced E6 transcripts in HPV16 + cell lines SCC090, SiHa, Ca Ski, in stable cell lines U-E6All #1, #6 and #7 and in HPV16 + HNSCC tumor samples. RPLP0 was used as loading control. Note that a band can be observed at 400 bp and could correspond to heteroduplexes between the different amplicons generated during PCR reaction. Full-length gels are presented in Figure . (E) RT-PCR showing alternatively spliced E6 transcripts in naturally HPV16-infected cells (W12_20861, W12_20862, W12_20863).
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    Expression profiles of HPV16 E6 and E6*I in stable cell lines. ( A ) Western blotting analysis showing p53, E6 and β-actin protein levels in U-2 OS parental cell line (U-CTL#par) and in stably transfected cells with pXJ40 empty vector (U-CTL #1 and #3) or pXJ40-E6All vector (U-E6All #1, #6 and #7) or pXJ40-E6*I vector (U-E6*I #1, #6 and #14). β-actin is used as loading control. Full-length blots are presented in Figure . ( B ) Western blotting analysis comparing protein levels of E6 and p53 between U-2 OS cellular clones (U-CTL#par, U-E6All #1, #6 and #7) and naturally HPV16-infected cells (W12_20861, W12_20862, W12_20863, SiHa and Ca Ski cell lines). β-actin is used as loading control. (C) RT-PCR analysis of alternatively spliced HPV16 E6 transcripts in U-E6All and U-E6*I stably transfected cells. As expected no E6 transcripts were observed in U-2 OS parental cell line (U-CTL#par) and in U-CTL #1 and #3 stably transfected cells. The detection of the vector in stably transfected cells was done by pXJ40 vector specific primers and RPLP0 was used as loading control. Full-length gels are presented in Figure . (D) RT-PCR analysis of alternatively spliced E6 transcripts in HPV16 + cell lines SCC090, SiHa, Ca Ski, in stable cell lines U-E6All #1, #6 and #7 and in HPV16 + HNSCC tumor samples. RPLP0 was used as loading control. Note that a band can be observed at 400 bp and could correspond to heteroduplexes between the different amplicons generated during PCR reaction. Full-length gels are presented in Figure . (E) RT-PCR showing alternatively spliced E6 transcripts in naturally HPV16-infected cells (W12_20861, W12_20862, W12_20863).

    Journal: Scientific Reports

    Article Title: Comparative RNA sequencing reveals that HPV16 E6 abrogates the effect of E6*I on ROS metabolism

    doi: 10.1038/s41598-019-42393-6

    Figure Lengend Snippet: Expression profiles of HPV16 E6 and E6*I in stable cell lines. ( A ) Western blotting analysis showing p53, E6 and β-actin protein levels in U-2 OS parental cell line (U-CTL#par) and in stably transfected cells with pXJ40 empty vector (U-CTL #1 and #3) or pXJ40-E6All vector (U-E6All #1, #6 and #7) or pXJ40-E6*I vector (U-E6*I #1, #6 and #14). β-actin is used as loading control. Full-length blots are presented in Figure . ( B ) Western blotting analysis comparing protein levels of E6 and p53 between U-2 OS cellular clones (U-CTL#par, U-E6All #1, #6 and #7) and naturally HPV16-infected cells (W12_20861, W12_20862, W12_20863, SiHa and Ca Ski cell lines). β-actin is used as loading control. (C) RT-PCR analysis of alternatively spliced HPV16 E6 transcripts in U-E6All and U-E6*I stably transfected cells. As expected no E6 transcripts were observed in U-2 OS parental cell line (U-CTL#par) and in U-CTL #1 and #3 stably transfected cells. The detection of the vector in stably transfected cells was done by pXJ40 vector specific primers and RPLP0 was used as loading control. Full-length gels are presented in Figure . (D) RT-PCR analysis of alternatively spliced E6 transcripts in HPV16 + cell lines SCC090, SiHa, Ca Ski, in stable cell lines U-E6All #1, #6 and #7 and in HPV16 + HNSCC tumor samples. RPLP0 was used as loading control. Note that a band can be observed at 400 bp and could correspond to heteroduplexes between the different amplicons generated during PCR reaction. Full-length gels are presented in Figure . (E) RT-PCR showing alternatively spliced E6 transcripts in naturally HPV16-infected cells (W12_20861, W12_20862, W12_20863).

    Article Snippet: Human osteosarcoma derived cell line U-2 OS, HPV16-positive carcinoma derived cell lines SiHa (ATCC® HTB-35™), SCC090 (ATCC® CRL-3239™) and U-2 OS stable cell lines were cultured in DMEM medium (Lonza, Verviers, Belgium).

    Techniques: Expressing, Stable Transfection, Western Blot, Transfection, Plasmid Preparation, Clone Assay, Infection, Reverse Transcription Polymerase Chain Reaction, Generated