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. 2020 Jul 8;15(7):e0235766. doi: 10.1371/journal.pone.0235766

Fig 4.

Fig 4

A: JAK3 expression in Ba/F3 JA K3 mutant cell lines transduced with JAK3 or control mutant vectors after selection in vitro with IL-3. JAK3 expression was assessed by western blot in Ba/F3 cell lines (selected in vitro with IL-3) with specific anti-JAK3 (N-term and C-term) antibodies and anti-GADPH antibody. B-M: Cell proliferation of mutant and wild-type cell lines upon IL-3 withdrawal assessed using the IncuCyte® ZOOM System. Cells were resuspended in 10% FCS RPMI medium with (plus_IL3) and without IL3 (minus_IL3) in a 96-well plate format and followed for 13 days. The x-axis reports time in days, the y-axis reports cell confluence in percentage (%). JAK3(N) = N-terminal antibody; JAK3(C) = C-terminal antibody; GAPDH = glyceraldehyde-3-phosphate dehydrogenase antibody; p.L1091P = JAK3 mutant #1; p.T1022I = JAK3 mutant #2; p.G888A = JAK3 mutant #3; JAK3 wt = JAK3 wild-type; pMSCV empty = pMSCV empty vector; BRAFV600E = pBabe-Puro-BRAF-V600E mutant; MEK-DD = pBabe-Puro-MEK-DD mutant; Myr-AKT = pBabe-Puro-Myr-AKT mutant; pBP empty = pBabe-Puro empty vector; BA/F3 = wild-type Ba/F3 cell lines; IL-3 = Interleukin 3.