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. 2000 Aug 15;106(4):501–509. doi: 10.1172/JCI9148

Figure 4.

Figure 4

Regulation of leptin-receptor expression during wound healing in Balb/c (wt) and C57BL/6J-ob/ob mice. (a) C57BL/6J mice were intraperitoneally treated with leptin as described in Methods. Total cellular RNA (20 μg) from nonwounded and wounded back skin of C57BL/6J-ob/ob mice treated with leptin as indicated was analyzed by RNase protection assay with an RNA hybridization probe complementary to ObRb and ObRa leptin–receptor splice variants. For every experimental time point, three wounds each from three animals (total: n = 9 wounds) were pooled for analysis. The time after injury is indicated for each lane. Control skin refers to nonwounded skin. Hybridization probe (1000 counts/min) was added to the lane labeled probe. Expression of GAPDH mRNA is shown as a loading control in the bottom panel. (b) Total protein (50 μg) from lysates of nonwounded and wounded back skin (day 1, 3, 5, 7, and 13 after injury, indicated for each lane) of C57BL/6J-ob/ob mice treated with leptin or PBS (used as control) as indicated were analyzed by immunoblotting for the presence of leptin-receptor subtype proteins ObRa and ObRb. Two wounds from the backs of three animals (n = 6) were excised for each experimental time point and used for protein isolation. Leptin-receptor subtypes ObRa and ObRb are indicated by arrowheads.