ATCC (Manassas, VA) provided HK-2 human renal proximal tubule cells. An HK2 cell line was grown in a keratinocyte serum-free medium (Invitrogen) and tested at generations 10-13. Following pretreatment with/without anlotinib (2 μM) for 4 h, cells were incubated with TGF-β1 for 48 h. Thereafter, cell lysates and supernatants were prepared as described below.
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