Colony-Formation Assay for Radiation Sensitivity

The colony-formation assay was performed as described previously [17 (link), 18 (link)] Colonies were fixed and stained with methylene blue solution (0.25% methylene blue (FUJIFILM Wako) in 90% ethanol (FUJIFILM Wako). The number of surviving colonies that included 50 cells or more was counted. The surviving fraction was calculated based on the plating efficiency [19 (link)]. The sensitivity enhancement ratio at 6 Gy (SER) was calculated using the equation SER₍₆₎ = log Sf_{(MHY1485, 6)} / log Sf_{(DMSO, 6)}, where Sf_{(MHY1485, 6)} and Sf_{(DMSO, 6)} are the surviving fractions at 6 Gy following exposure to MHY1485 and DMSO, respectively [20 (link)].

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Sun L., Morikawa K., Sogo Y, & Sugiura Y. (2021). MHY1485 enhances X-irradiation-induced apoptosis and senescence in tumor cells. Journal of Radiation Research, 62(5), 782-792.

Publication 2021

methylene blue ethanol

Assay Cells Dmso Ethanol Methylene blue Mhy1485 Sensitivity

Corresponding Organization : National Institute of Advanced Industrial Science and Technology

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Variable analysis

independent variables

Exposure to MHY1485

dependent variables

Number of surviving colonies with 50 or more cells
Surviving fraction
Sensitivity enhancement ratio at 6 Gy (SER)

control variables

Exposure to DMSO

controls

Positive control: Exposure to DMSO
Negative control: Not explicitly mentioned

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