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. 2018 Sep 26;14(9):e1007679. doi: 10.1371/journal.pgen.1007679

Fig 2. Rapamycin has similar effects in growth inhibition of RCC 704 cells independent of PBRM1 addback.

Fig 2

(A) Immunoblot analysis of the PBRM1 null A704 cell line stably transfected with doxycycline inducible constructs expressing empty vector (EV), wild-type PBRM1 (WT) or mutant Q1298* PBRM1, treated with or without doxycycline (Dox, 1ug/ml) for 5 days. (B) Growth of these same A704 derivative cell lines treated with or without doxycycline and with DMSO (control) or rapamycin (RAPA, 20nM) for 2, 4, or 6 days. Cell growth was assessed by Crystal Violet staining. (C) Proportional growth of A704 derivative cell lines derived from panel B. Rapamycin inhibited the growth of all 3 cell lines to a similar extent, and doxycycline treatment had little or no effect on growth. (D, E) These same A704 derivative cell lines were plated at 200 cells per 10cm dish and treated with or without Doxycycline and DMSO control or Rapamycin (20nM) for 30 days, then formalin fixed and stained with Crystal Violet. D, Image of colonies after crystal violet staining. E, Quantitation of colonies > 2mm in diameter per 10 cm dish. The bar is placed at the median value.