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. 2020 Feb 7;9(4):145–160. doi: 10.1089/wound.2019.0956

Figure 2.

Figure 2.

hsa_circ_0084443 localizes in the cytoplasm of human epidermal keratinocytes and its aberrant expression does not change PRKDC levels. (A) Northern blotting analysis of hsa_circ_0084443 expression in human primary keratinocytes (HEKa) (lane 2, 3) and skin tissue (lane 4–7) with and without RNase R treatment. RNA molecular weight marker was used (lane 1). (B) qRT-PCR analysis of hsa_circ_0084443 and PRKDC mRNA in HEKa treated with Actinomycin D at the indicated timepoints (n = 3). (C) qRT-PCR analysis of hsa_circ_0084443, MALAT1, and GAPDH in nucleus or cytoplasm of HEKa (n = 3). (D) Fluorescence in situ hybridization to visualize hsa_circ_0084443 in HEKa. Scale bar 20 μm. (E) Illustration of hsa_circ_0084443 and pre-mRNA of PRKDC. Sequence information of si-hsa_circ_0084443, si-control, and si-hsa_circ_0084443-control. qRT-PCR of hsa_circ_0084443 (F) and PRKDC mRNA (H) in keratinocytes transfected with 10, 20, and 50 nM si-control, si-hsa_circ_0084443, or si-hsa_circ_0084443-control for 24 h (n = 3). QRT-PCR of hsa_circ_0084443 (G) and PRKDC mRNA (I) in keratinocytes transfected with hsa_circ_0084443 overexpression plasmid (0.1 or 0.2 μg) for 24 h (n = 3) **p < 0.01 and ***p < 0.001 by unpaired two-tailed Student's t test. Data are presented as mean + SD. Color images are available online.