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. 2011 Feb 16;300(5):C1139–C1154. doi: 10.1152/ajpcell.00167.2010

Fig. 3.

Fig. 3.

Pnck expression suppresses the level of the ATP-independent small heat shock chaperone Hsp27; however, this suppression does not explain the Pnck-induced suppression of EGFR levels. A: Hsp27 levels are suppressed by Pnck. Stable Neo (lane 1) and HA-Pnck (lane 2) HEK-293 cells were plated at low cell density and allowed to grow for 48 h. Cells were lysed, and equal amounts of protein were immunoblotted for EGFR, HA-Pnck, β-actin, Hsp90, Hsp70, Hsc70, Hop (p60), p23, and Hsp27 expression. The experiment was conducted thrice with essentially identical results. B: short hairpin RNA (shRNA)-mediated knockdown of endogenous Hsp27 expression does not result in suppression of EGFR levels. Parental HEK-293 cells were infected with either control virus (lane 1) or one of several Hsp27 shRNA lentiviruses (lanes 2–6) in complete medium using polybrene. Medium was removed after 24 h, and the cells were supplemented with fresh medium. Cells were lysed after an additional 24 h, and equal amounts of total lysate were immunoblotted for EGFR (WB: EGFR), Hsp27 (WB: Hsp27), and Hsp90 (WB: Hsp90) expression.