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. Author manuscript; available in PMC: 2014 Sep 19.
Published in final edited form as: J Invest Dermatol. 2012 Feb 16;132(5):1435–1442. doi: 10.1038/jid.2012.14

Figure 5. Normal human epidermal keratinocytes (NHEKs) pretreated with doxycycline display inhibited proteolytic processing of recombinant hCAP18.

Figure 5

Equal amounts of recombinant cathelicidin precursor (rhCAP18, 1.525 µg) were added as indicated to identical cultures of live NHEKs either untreated or pretreated overnight with doxycycline (225 µm). Cleavage of hCAP18 was monitored by western blot analysis with anti-LL-37 antibody. (a) Cathelicidin detected in whole-cell extract and (b) cathelicidin remaining in overlying NHEK culture media. Lane 1, rhCAP18 alone + NHEK; lane 2, NHEK alone; lane 3, rhCAP18 + doxycycline + NHEK; lane 4, doxycycline alone + NHEK. Anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is shown as loading control. In the absence of doxycycline (lane 1), rhCAP18 is enzymatically processed as detected by generation of a band at the expected migration of LL-37. In the presence of doxycycline (lane 3), LL-37 is not seen in the cell-associated fraction, and a greater amount of rhCAP18 remains in the supernatant.