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Fig. 2 | Journal for ImmunoTherapy of Cancer

Fig. 2

From: Therapeutic efficacy of a novel humanized antibody-drug conjugate recognizing plexin-semaphorin-integrin domain in the RON receptor for targeted cancer therapy

Fig. 2

H5B14-induced cell surface RON internalization: Four cancer cell lines HT-29, L3.6pl, H358, and T-47D at 1 × 106 cells per dish were treated at 37 °C with 5 μg/ml of H5B14 or PCM5B14, collected at different time points, washed with acidic buffer to eliminate cell surface bound IgG [1], and then incubated with 2 μg/mL of anti-RON mAb Zt/c1 [23]. Immunofluorescence was analyzed by flow cytometer using FITC-coupled anti-mouse IgG. Immunofluorescence from cells treated with H5B14 at 4 °C was set as 100%. Internalization efficiency (IC50) was calculated as the time required to achieve a 50% reduction of cell surface RON

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