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. 2017 Jul 31;12(7):e0182495. doi: 10.1371/journal.pone.0182495

Fig 8. Cx46, Cx50 and Cx43 expression in wild type mice and in leptin receptor- (db/db) and leptin- (ob/ob) deficient male mice.

Fig 8

Wild type (WT), leptin receptor- (db/db) and leptin- (ob/ob) deficient male mouse anterior pituitaries were harvested. Thirty μg of total protein samples were loaded per well. After electrophoretic migration, proteins were electrotransferred onto nitrocellulose membranes and probed with either Cx46, Cx50, Cx43 or Prl antibodies. Membranes were reprobed with monoclonal or polyclonal anti-actin. Bands were scanned and their intensity quantified. The values obtained were normalized to the corresponding actin band intensity value. The figure shows representative Western blots. The values are the mean ± SEM of three different animals per experimental condition. Statistics (Student’s t test): Cx46: Total Cx46 levels were not significantly different in db/db and in WT mice. However, the intensities of the 14 and 48–49 kDa Cx46 immunoreactive bands were lower in db/db than in WT mice (+ P<0.01 and # P<0.02 respectively) whereas that of 68 kDa was stronger db/db than WT mice (+ P<0.01). Total, 14, 48–49 and 68 kDa Cx46 levels were significantly decreased (* P<0.05, + P<0.01, ** P<0.005 and † P<0.03 respectively) in ob/ob compared to WT mice. Cx50: Total Cx50 levels showed little change in db/db and ob/ob mice compared to WT. The intensity of the 51 kDa Cx50 immunoreactive band was more intense in db/db and ob/ob mice compared to WT mice (+ P<0.01). By contrast, the intensity of the 61 kDa-immunoreactive band was weaker in db/db and ob/ob mice compared to WT mice (* P<0.05 and +P<0.01 respectively). Cx43: Cx43 total levels (P0+P1+P2) increased in db/db compared to WT mice (+ P<0.01) due to the increase in the phosphorylated P2 band (++ P<0.001). P0 and P1 Cx43 showed no significant differences in db/db and WT mice. However, all Cx43 isoforms decreased in ob/ob compared to WT mice (* P<0.05). Prl: Anterior pituitary Prl content decreased (* P<0.05) in db/db but increased (* P<0.05) in ob/ob mice compared to the WT counterparts.