Skip to main content
. 2015 Mar 23;290(19):11918–11934. doi: 10.1074/jbc.M115.645903

FIGURE 6.

FIGURE 6.

SMAD3 gene silencing reduces the formation of p-SMAD3 (Ser-213)-containing complexes and suppresses iron-induced C3 up-regulation. A, Western blot and densitometry analysis for SMAD3, p-SMAD3 (Ser-213), and TFRC of ARPE-19 cell lysates derived from untransduced (control), vector, sh-null, and sh-SMAD3 lines, each in the absence or presence of FAC. Efficient knockdown of SMAD3 decreased the formation of p-SMAD3 (Ser-213)-containing complexes at 150 kDa. B, the FAC-induced increase in C3 mRNA levels is observed in the untransduced, vector, and sh-null cell lines but is absent in the sh-SMAD3 cell line. C, Western blot for non-nuclear (Non-Nucl) and nuclear (Nucl) lysates of untreated and FAC-treated cells. The p-SMAD3 (Ser-213)-containing complex at 150 kDa localizes only to the non-nuclear fractions. The p-SMAD3 (Ser-213) antibody-detectable band at 52 kDa is essentially unchanged by FAC treatment in both the non-nuclear and nuclear fractions. Data are expressed as mean ± S.E. (error bars) (n ≥ 3 with the following statistical notations: *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001; ns, not significant). d, days.