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

Figure 6.

Figure 6

H11/HspB8 upregulates Beclin-1 through TAK1/pmTORS2481. (a) Serial sections from A2058 and A375 xenografts as in Figure 1b were stained by TUNEL (TMR-red labeled dUTP) and with antibody to Beclin-1 (AlexaFluor 488-conjugated secondary antibody; green) and the percentage of cells with co-localized staining was calculated relative to the total cell number determined by DAPI staining. (b) Extracts from stably transfected A375 and A2058 cells given Dox (5 μg/ml; 3 days) in the absence or presence of the TAK1 dominant-negative mutant K63W or empty vector were immunoblotted with antibodies to Beclin-1 or actin. Data were quantified by densitometric scanning and the results are expressed as Beclin-1/actin densitometric units±S.D. (c) Extracts from cell cultures that were untreated or Dox treated in the presence or absence of K63W were immunoblotted with antibodies to pmTORS2481, pmTORS2448 or actin. Blots were stripped between probings. Data were quantified by densitometric scanning and the results are expressed as p-mTOR/actin densitometric units±S.D. (d) Extracts from stably transfected A2058 cells treated or not with Dox (5 μg/ml) in the presence or absence of rapamycin (20 nM) were immunoblotted with Beclin-1 followed by actin antibodies. Data were quantified by densitometric scanning and the results are expressed as Beclin-1/actin densitometric units±S.D.