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. Author manuscript; available in PMC: 2011 Nov 1.

Published in final edited form as: Cancer Biol Ther. 2010 Apr 1;9(7):526–536. doi: 10.4161/cbt.9.7.11116

Figure 4A and B. MDA-7/IL-24 stimulates expression of MDA-7/IL-24 in GBM cells in a PERK-dependent fashion. (A) GBM6 cells were infected with Ad.cmv or Ad.mda-7. Forty-eight h after infection cells and media were isolated. Cell viability was determined by trypan blue dye exclusion assay (±SEM, n = 3). The media was placed onto GBM6 cells and 24 h after media addition cells were irradiated (4 Gy). Forty-eight h after placement of conditioned media onto the cells, cell viability was again determined by trypan blue dye exclusion assay (±SEM, n = 3). (B) GBM6 cells were infected with Ad.cmv or Ad.mda-7. Forty-eight h after infection cells and media were isolated. Cell viability was determined by trypan blue dye exclusion assay (±SEM, n = 3). The media was placed onto GBM6 cells that had been transfected with plasmids to express a control shRNA (shSCR) or an shRNA to knock down MDA-7/IL-24 expression (shMDA-7) (labeled in parentheses, grey). Thirty min after media addition cells were treated with: (a) radiation (4 Gy) or; (b) vehicle (DMSO) or OSU-03012 (OSU, 1 μM). Forty-eight h after placement of conditioned media onto the cells, cell viability was again determined by trypan blue exclusion assay (±SEM, n = 3). Inset Panel: conditioned media containing MDA-7/IL-24 induces MDA-7/IL-24 in uninfected GBM cells that is blocked by shRNA knock down of MDA-7/IL-24.

Figure 4C and D. MDA-7/IL-24 stimulates expression of MDA-7/IL-24 in GBM cells in a PERK-dependent fashion. (C) GBM6 cells were infected with Ad.cmv or Ad.mda-7. Forty-eight h after infection cells and media were isolated. Cell viability was determined by trypan blue exclusion assay (±SEM, n = 3). The media was placed onto GBM6 cells, Twenty-four h after media addition cells were treated with: vehicle (DMSO) or OSU-03012 (OSU, 1 μM). Forty-eight h after placement of conditioned media onto the cells, cell viability was again determined by trypan blue dye exclusion assay (±SEM, n = 3). (D) GBM6 cells were infected with Ad.cmv or Ad.mda-7. Forty-eight h after infection the growth media was isolated. The media was placed onto GBM6 cells that had been transfected with plasmids to express a nothing (CMV) or to express dominant negative PERK (dnPERK) (labeled in parentheses, grey). Forty-eight h after placement of conditioned media onto the cells, cell viability was again determined by trypan blue exclusion assay (±SEM, n = 3) *p < 0.05 less than corresponding value in parallel empty vector control transfected cells.

Figure 4A and B. MDA-7/IL-24 stimulates expression of MDA-7/IL-24 in GBM cells in a PERK-dependent fashion. (A) GBM6 cells were infected with Ad.cmv or Ad.mda-7. Forty-eight h after infection cells and media were isolated. Cell viability was determined by trypan blue dye exclusion assay (±SEM, n = 3). The media was placed onto GBM6 cells and 24 h after media addition cells were irradiated (4 Gy). Forty-eight h after placement of conditioned media onto the cells, cell viability was again determined by trypan blue dye exclusion assay (±SEM, n = 3). (B) GBM6 cells were infected with Ad.cmv or Ad.mda-7. Forty-eight h after infection cells and media were isolated. Cell viability was determined by trypan blue dye exclusion assay (±SEM, n = 3). The media was placed onto GBM6 cells that had been transfected with plasmids to express a control shRNA (shSCR) or an shRNA to knock down MDA-7/IL-24 expression (shMDA-7) (labeled in parentheses, grey). Thirty min after media addition cells were treated with: (a) radiation (4 Gy) or; (b) vehicle (DMSO) or OSU-03012 (OSU, 1 μM). Forty-eight h after placement of conditioned media onto the cells, cell viability was again determined by trypan blue exclusion assay (±SEM, n = 3). Inset Panel: conditioned media containing MDA-7/IL-24 induces MDA-7/IL-24 in uninfected GBM cells that is blocked by shRNA knock down of MDA-7/IL-24.

Figure 4C and D. MDA-7/IL-24 stimulates expression of MDA-7/IL-24 in GBM cells in a PERK-dependent fashion. (C) GBM6 cells were infected with Ad.cmv or Ad.mda-7. Forty-eight h after infection cells and media were isolated. Cell viability was determined by trypan blue exclusion assay (±SEM, n = 3). The media was placed onto GBM6 cells, Twenty-four h after media addition cells were treated with: vehicle (DMSO) or OSU-03012 (OSU, 1 μM). Forty-eight h after placement of conditioned media onto the cells, cell viability was again determined by trypan blue dye exclusion assay (±SEM, n = 3). (D) GBM6 cells were infected with Ad.cmv or Ad.mda-7. Forty-eight h after infection the growth media was isolated. The media was placed onto GBM6 cells that had been transfected with plasmids to express a nothing (CMV) or to express dominant negative PERK (dnPERK) (labeled in parentheses, grey). Forty-eight h after placement of conditioned media onto the cells, cell viability was again determined by trypan blue exclusion assay (±SEM, n = 3) *p < 0.05 less than corresponding value in parallel empty vector control transfected cells.