IGF-1 treatment attenuates anti-cancer efficiency of zoledronic acid in HeLa cells derived CSCs. Western blot analysis of phosphorylated Erk1/2 and Akt after treatment with IGF-1 for 6, 12 and 24 h in HeLa cells derived CSCs, respectively (a). Control vs. 6, 12, and 24 h treatment with IGF-1: * P < 0.05, ** P < 0.01. Western blot analysis of stemness-, EMT-, apoptosis-, and cell cycle-associated proteins in HeLa cells derived CSCs treated or not with IGF-1 and zoledronic acid (ZOL) (b-d). DAPI staining of apoptotic cervical CSCs after being treated or not with IGF-1 and ZOL. Typical apoptotic bodies in HeLa cells derived CSCs are shown with red arrows. The histogram shows the proportions of DAPI-stained apoptotic HeLa cells derived CSCs after being treated or not with IGF-1 and ZOL (e). The migrated HeLa cells derived CSCs after being treated or not with IGF-1 and ZOL. The histogram shows the number of migrated HeLa cells derived CSCs; original magnification, × 200 (f). The histogram shows the SFE of HeLa cells derived CSCs treated or not with IGF-1 and ZOL, respectively (g). The histogram shows the proportions of cell cycle distribution in G1, S, and G2/M phases after being treated or not with IGF-1 and ZOL, respectively (h). ZOL(−) plus IGF-1(−) vs. ZOL(+) plus IGF-1(−) or ZOL(−) plus IGF-1(+): * P < 0.05, ** P < 0.01. ZOL(+) plus IGF-1(−) vs. ZOL(+) plus IGF-1(+): # P < 0.05, ## P < 0.01. Scale bars represent 50 μM in inset. Results are shown as mean values ± SD of independent experiments performed in triplicate