A Set of Proximal Regulatory Elements Contribute to the Transcriptional Activity of the Human Lipoprotein Lipase Promoter

Figures

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  Figure 1. Overview: A schematic representation of the methods used for this study (created with Biorender.com) on 14 October 2024. Refer to Supplementary Figure S1 for the colony PCR.

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  Figure 2. The 12 key regulatory motifs in the 5′ promoter region of the Homo sapiens LPL gene. The numbering is relative to the first nucleotide of the first exon (+1). The primers used to amplify the full promoter sequence are underlined and highlighted in yellow and the primers used to amplify the partial promoter sequence are underlined and highlighted in dark gray. The capital letters represent the amino acids encoded by exon 1 (highlighted in red) (Sequence was adapted from the human genome assembly reference sequence NG_008855.2).

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  Figure 3. Agarose gel showing the resolved plasmids harboring full (a) or partial (b) LPL promoters cloned and inserted into the pGL4.10[luc2] luciferase vector. The plasmids were digested with SacI and XhoI. The fragments were resolved on a 1% agarose gel prestained with 10 mg/mL SYBR Safe and imaged under UV light.

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  Figure 4. (a) A plasmid map of the promoterless pGL4.10[luc2] luciferase vector showing the regions amplified from the vector (modified from Promega; the red labels on the map indicate the primer regions). Agarose gel showing the full and partial promoter inserts amplified from the recombinant luciferase vectors using MCS primers in (b) and luciferase-specific primers in (c).

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  Figure 5. A comparison of luciferase activity obtained using the different tested vectors to determine the effect of the full and partial LPL promoters on pGL4.10 gene expression in HEK-293 cells. This effect was measured as the mean of the firefly/Renilla luciferase ratio. The results are presented as the means from three independent experiments conducted in triplicate (* p ≤ 0.05, ** p ≤ 0.01).