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Fig. 3 | BMC Research Notes

Fig. 3

From: ER stress regulates alkaline phosphatase gene expression in vascular smooth muscle cells via an ATF4-dependent mechanism

Fig. 3

ATF4 does not bind the ALP promoter in VSMCs. a ER stress-related transcription factor binding sites mapped to the ALP promoter. b Schematic of ALP promoter constructs with potential regions activating (red) and inhibiting (blue) transcription in VSCMs, based on luciferase assay results. ‘+’ denotes degrees of activation, ‘−’ denotes lack of promoter activity. c The fragment of the ALP promoter containing potential ATF4 binding sites was amplified using biotinylated primers and conjugated onto magnetic streptavidin beads. The beads were then incubated with VSMC nuclear extracts, washed and proteins were eluted off the DNA and analysed by Western blotting. Coomassie gel demonstrates that even though ATF4 was not detected in the pulldown sample, other proteins were present. N = 3, figure shows representative images

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