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. 2018 May 4;9(34):23334–23348. doi: 10.18632/oncotarget.25101

Figure 3. PTPRJ overexpression dephosphorylates CD98hc and downregulates its protein levels.

Figure 3

(A, B) A549 cells were seeded in 100 mm culture dishes and, after twenty-four hours, transduced by a recombinant Ad PTPRJ at MOI50. Forty-eight hours later, cells were lysed and extracted proteins loaded on polyacrylamide gel, transferred to nitrocellulose filter, and analyzed by Western blot through CD98hc, PTPRJ and p-tyrosine antibodies. γ-tubulin was used to normalize protein loading. (C) An immunofluorescence was also performed. A549 cells were transduced with Ad PTPRJ at MOI50. Forty-eight hours later, A549 cells were incubated with PTPRJ mAb and CD98hc mAb; afterwards, secondary rabbit-633 Ab (green) and mouse-PE Ab (red) were added to stain cells, as described in Materials and Methods, and observed by confocal microscopy. CD98hc levels decrease with PTPRJ overexpression (white arrow) in contrast with a lower expression of PTPRJ where CD98hc levels do not change (yellow arrow).