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. 2024 Apr 22;20(7):2640–2657. doi: 10.7150/ijbs.95595

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Restoring FKBP12 partially reverses the effects of HMGA1 depletion on cell sensitivity to rapamycin. (A) KYSE510 shRNA ctrl and shRNA-HMGA1 (control vs shHMGA1) cells were transduced with pcDNA3.1/FKBP12 and treated with 1 μM rapamycin. Whole cell extracts were isolated for the detection of HMGA1, FKBP12, mTOR downstream effectors, and beta-actin (loading control) with western blot. (B) TE13 empty vector and HMGA1 overexpression (vector vs oeHMGA1) cells were transduced with FKBP12-shRNA and treated with 1 μM rapamycin. Whole cell extracts were isolated for the detection of HMGA1, FKBP12, mTOR downstream effectors, and beta-actin (loading control) with western blot. (C) KYSE510 shRNA ctrl and shRNA-HMGA1 (control vs shHMGA1) cells were transduced with pcDNA3.1/FKBP12 and treated with 1 μM rapamycin. Colony formation assay was performed in the cells. Representative images of colony formation are shown in the upper panel. Colonies with 50 or more cells were counted. Calculations of relative colonies in each treatment (combination of treatments) are shown in the lower panel. (D) TE13 empty vector and HMGA1 overexpression (vector vs oeHMGA1) cells were transduced with FKBP12-shRNA and treated with 1 μM rapamycin. Colony formation assay was performed in the cells. Representative images of colony formation are shown in the upper panel. Colonies with 50 or more cells were counted. Calculations of relative colonies in each treatment (combination of treatments) are shown in the lower panel.