Fig. 4
From: Cooperative antitumor activities of carnosic acid and Trastuzumab in ERBB2+ breast cancer cells
CA and Tz cooperatively upregulate p27KIP1 expression levels in SKBR-3 and BT474 cells. Cells were cultured for 7d with control medium (Ctr), CA, Tz or CA + Tz supplemented medium, which was changed every other day. a A representative immunoblot analysis is shown, which was performed with anti-p27KIP1 antibody on whole cell lysates. Tubulin is shown as loading control. Numbers on each lane represent p27KIP1 protein levels determined as described in Methods. b Epifluorescence images of SKBR-3 and BT474 cells cultured for 7d with CA + Tz fixed, permeabilized and challenged with anti p27KIP1 antibody and Alexa488-conjugated anti-mouse or with Alexa488-conjugated anti-mouse only as negative secondary antibody control (green signal). Nuclei were stained with DAPI (red signal). The extent of overlap is shown in yellow in the merged images (rightmost column) and arrowheads indicate some cells displaying p27KIP1 nuclear localization. Bar = 20 μm