Genetically Engineered PDAC Cell Line

KPsh cells (kindly donated by Scott W Lowe) derive from PDAC that developed in a Pdx1-cre;LSL-Kras^G12D;Col1a1-TRE-shp53-shRenilla;Rosa26-CAGs-LSL-rtTA-IRES-mKate2 mice [25 (link)]. Cells were grown in DMEM (10% FBS Gibco; penicillin–streptomycin 2 units/ml) at 37 °C, 5% CO₂ and they were propagated on collagen-coated plates (PurCol, Advanced Biomatrix, 0.1 mg/ml). KPsh were maintained in 1 μg/ml doxycycline to keep off p53 through doxycycline-dependent shRNA against Trp53; to allow p53 expression the doxycycline was removed 48 h before further procedure. KPsh cells have been authenticated as described in the source papers (references above); we also confirm p53 status by RT-qPCR or western blot.

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Panatta E., Butera A., Celardo I., Leist M., Melino G, & Amelio I. (2022). p53 regulates expression of nuclear envelope components in cancer cells. Biology Direct, 17, 38.

Publication 2022

PurCol

Cells Collagen Doxycycline Ires Mice Pdac Pdx1 Penicillin Shrna Streptomycin Trp53 Western blot

Corresponding Organization :

Other organizations : University of Rome Tor Vergata, University of Konstanz

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Variable analysis

independent variables

Presence of doxycycline (1 μg/ml) to keep p53 shRNA off

dependent variables

P53 expression (measured by RT-qPCR or western blot)

control variables

Cell culture media (DMEM with 10% FBS and penicillin-streptomycin)
Incubation conditions (37°C, 5% CO2)
Cell propagation on collagen-coated plates (0.1 mg/ml)

controls

Positive control: KPsh cells maintained in 1 μg/ml doxycycline to keep p53 shRNA on and suppress p53 expression
Negative control: KPsh cells with doxycycline removed 48 h before further procedure to allow p53 expression

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