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. 2016 Mar 24;7:358. doi: 10.3389/fpls.2016.00358

Figure 2.

Figure 2

hBMP2nat and zein-hBMP2ad are soluble in transiently transfected protoplasts and are not secreted. (A,B) Tobacco leaf protoplasts were transiently transfected with plasmid encoding hBMP2nat (lanes 5–6, 11–12), zein-hBMP2ad (lanes 3–4, 9–10), or empty plasmid lanes (1–2, 7–8) and incubated for 24 h. (A) Protoplasts or incubation media were homogenized in the presence (+) or absence (−) of 2-ME. Aliquots corresponding to 20.000 protoplasts (cells) or the corresponding incubation medium (medium) were analyzed by SDS-PAGE in reducing conditions, followed by protein blot with anti-Flag antiserum. (B) Aliquots corresponding to 16.500 protoplasts were homogenized in the presence (+) or absence (−) of 2-ME. After centrifugation, soluble (S) material and insoluble precipitate (I) were analyzed by SDS-PAGE in reducing conditions, followed by protein blot with anti-Flag antiserum. Asterisk: 18 kD fragment of hBMP2ad. Numbers on the left indicate the positions of molecular mass markers, in kD.