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. 2012 Aug 16;3(8):e371. doi: 10.1038/cddis.2012.108

Figure 1.

Figure 1

Restored H11/HspB8 expression inhibits the growth of A375 and A2058 xenografts. (a) Untransfected A375 and A2058 cells were treated with Dox (5 μg/ml; 3 days) and stained in triple immunofluorescence with antibodies to S100 (AlexaFluor350-conjugated secondary antibody; blue) and H11/HspB8 (AlexaFluor 488-conjugated secondary antibody; green) and TUNEL (TMR-red labeled dUTP). The cells were counted in five randomly selected 3 mm2 fields (250 cells each) and the % positive cells calculated relative to the total number of melanoma cells (S100+). Results are expressed as % positive cells±S.D. Similar results were obtained for Dox at 2 or 10 μg/ml. (b) Balb/c (nu/nu) mice given A2058 or A375 cells stably transfected with tet-regulated H11/HspB8 (5 × 106 cells) by subcutaneous injection into both flanks were randomly divided into two groups (n=10 tumors/group) and left untreated or given Dox (2 mg/ml) in the drinking water. Tumor volume calculated as described in the Materials and methods section is shown for A2058 xenografts. Similar results were obtained for A375 xenografts (*P<0.001). Mice bearing A375 xenografts that were untreated (control) or treated with Dox (Dox) were photographed on day 25 and representative excised tumors are shown. (c) Mice given 5 × 105 stably transfected cells were similarly followed and tumor volume calculated (*P<0.001)