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Fig. 1 | Cell Communication and Signaling

Fig. 1

From: PML-II regulates ERK and AKT signal activation and IFNα-induced cell death

Fig. 1

IFNα-induced cell death is PML-II dependent. A HeLa cells were stimulated with 1000 U/ml IFNα or not and PML isoform mRNA levels measured by SYBR-Green qPCR. The relative expression level of each PML isoform was quantified relative to β-actin. B 30–50% confluent HeLa cells were transfected with siPML-II, siPML or control siRNA for 48 h, and following 1000 U/ml IFNα or mock stimulation for 12 h, cells were collected and mRNA levels for PML-II and total PML were measured by SYBR-Green qPCR. The relative mRNA expression of each gene was quantified relative to GAPDH, and normalized to the level of unstimulated control siRNA-transfected cells. C HeLa cells, D HEK293 cells were transfected with PML-II or control siRNA for 48 h, then stimulated with 1000 U/ml IFNα at the described time points or 1 μg/ml poly(I:C) for 12 h. Total protein lysates were analyzed by western blotting for PML and PML-II, respectively. E Hela cells were transfected with either siPML-II or control siRNA for 48 h, and stimulated with 1000 U/ml IFNα for 12 h, then mRNA levels of PML-I, PML-II, PML-IV and PML-V were measured by SYBR-Green qPCR as in (B). F HeLa cells were treated with 1000 U/ml IFNα for 24 h or 48 h, and cell death quantified by PI staining and flow cytometry. G Hela cells were transfected with either siPML-II or control siRNA for 24 h, and following stimulation for 24 h with amounts of IFNα as indicated, cell death was quantified as in (F). H HeLa cells were transfected with either siPML-II or control siRNA for 12 h, then following 1000 U/ml IFNα stimulation for 48 h, cell death was detected as (F). I HeLa cells were transfected with the amounts of poly(I:C) indicated for 24 h, then cell death quantified as in (F). J HeLa cells were transfected with siRNA as in (F) for 24 h, then with 1 μg/ml poly(I:C) for 24 h and cell death quantified as in (F). Error bars show standard deviation among technical replicates

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