Characterization of TSC2-deficient Kidney Tumor Cell Lines

TSC2-deficient kidney tumor cell lines, LAM 621-101 (human, Research Resource Identifier [RRID]: CBCL_S897) (28 (link)), 105K (mouse) (65 (link)), and TMKOC (mouse) (66 (link)), were provided by Drs Jane Yu and Elisabeth Henske. TMKOC was originally generated by Dr Vera Krymskaya (67 (link)). HEK293E cell line (RRID: CVCL_6974), MCF7 (RRID: CVCL_0031), BT549 (RRID: CVCL_1092), and UMB1949 (RRID: CVCL_C471) were obtained from American Type Culture Collection. LAM 621-101, UMB1949, MCF7, BT549, TMKOC, 105K, and HEK293E cells were grown in Dulbecco's modified Eagle's medium (GIBCO) with 10% fetal bovine serum (FBS) (Sigma–Aldrich) at 37 °C with 5% CO₂. About 5 × 10⁶ cells counted by Multisizer 4e Coulter Counter (Beckman) were plated on a 60 mm plate and serum starved for 24 h unless otherwise indicated. Rapamycin (Calbiochem) dissolved in dimethyl sulfoxide (DMSO) was treated at the final concentration of 20 nM (LAM 621-101, UMB1949, MCF7, and BT549) or 100 nM (TMKOC and 105K). MAPK13-IN-1 (MAPK13 inhibitor; MedChemExpress) dissolved in DMSO was treated at the final concentration of 5 μM unless otherwise indicated.

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Kim J., Chun Y., Ramirez C.B., Hoffner L.A., Jung S., Jang K.H., Rubtsova V.I., Jang C, & Lee G. (2023). MAPK13 stabilization via m6A mRNA modification limits anticancer efficacy of rapamycin. The Journal of Biological Chemistry, 299(9), 105175.

Publication 2023

BT549 fetal bovine serum (FBS) Multisizer 4e Coulter Counter

Cell line Cells Dimethyl sulfoxide Fetal bovine serum Human Kidney Mcf7 Mouse Rapamycin Serum Tsc2 Tumor cell lines

Corresponding Organization : University of California, Irvine

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Variable analysis

independent variables

Rapamycin treatment (20 nM or 100 nM)
MAPK13-IN-1 treatment (5 μM)

dependent variables

Not explicitly mentioned

control variables

Cell lines: LAM 621-101, UMB1949, MCF7, BT549, TMKOC, 105K, HEK293E
Culture media: Dulbecco's modified Eagle's medium (GIBCO) with 10% fetal bovine serum (FBS) (Sigma–Aldrich)
Incubation conditions: 37 °C with 5% CO2
Serum starvation for 24 h before treatments

controls

Positive control: Not specified
Negative control: Untreated cells (DMSO vehicle control)

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