Figure 5.

hnRNP K does not inhibit HSF1 activation nor its nuclear translocalization. A and B, HEK293T cells were transfected with Flag empty vector, Flag-hnRNP K, and Flag-hnRNP K C132S (A), and HeLa cells were transfected with control and hnRNP K siRNA (B). The cells were heat shock–treated at 45 °C for 15 min and recovered at 37 °C for the indicated times. The cells were analyzed by Western blot analysis using anti-HSF1 antibody to detect heat shock–induced band shift and anti-Flag antibody to check overexpression of Flag-tagged proteins. high, active higher molecular mass HSF1; low, inactive lower molecular mass HSF1; asterisk, nonspecific band. C and D, HEK293T cells were transfected with Flag empty vector, Flag-hnRNP K, and Flag-hnRNP K C132S (C), and HeLa cells were transfected with control and hnRNP K siRNA (D). The cells were heat shock–treated at 45 °C for 15 min. The cells were fractionated into cytosolic and nuclear fractions and analyzed by Western blot analysis using anti-HSF1 to detect its localization, anti-Flag antibody to check overexpression of Flag-tagged proteins, and anti-PRX6 and lamin B as cytosolic and nuclear markers, respectively. HS, heat shock treatment; Cont, control; IB, immunoblot; C, cytosolic fraction; N, nuclear fraction.