Figure 2.

Aberrant expression of HMGA1 in ESCC cells leads to altered responsiveness to rapamycin. (A) Colony-forming units in KYSE510 cells with or without HMGA1 silencing (control vs shHMGA1). Cells were simultaneously treated with 1 μM rapamycin for 48 h. Representative images of colony formation are shown. (B) KYSE510 cells with or without HMGA1 knockdown were treated with rapamycin in a concentration gradient (starting at 8 nM and increasing fivefold each time). Cell viability was measured using the CCK8 assay. (C) Colony-forming units in TE13 cells with or without HMGA1 overexpression (vector vs oeHMGA1). Simultaneously, the cells were treated with 1 μM rapamycin for 48 h. Representative images of colony formation are shown. (D) TE13 cells with or without HMGA1 overexpression were treated with rapamycin in a concentration gradient (starting at 8 nM and increasing fivefold each time). Cell viability was measured using the CCK8 assay. (E) HMGA1, FKBP12, mTOR downstream effectors, and beta-actin (loading control) in KYSE510 cells transduced with shHMGA1 were detected with immunoblot. Extracts of ESCC cells were collected 24 h after 1 μM rapamycin treatment. (F) HMGA1, FKBP12, mTOR downstream effectors, and beta-actin (loading control) in TE13 cells transduced with lentivirus-expressed HMGA1 were detected with immunoblot. Extracts of ESCC cells were collected 24 h after 1 μM rapamycin treatment.