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. Author manuscript; available in PMC: 2019 Sep 1.
Published in final edited form as: Pain. 2018 Sep;159(9):1777–1789. doi: 10.1097/j.pain.0000000000001271

Fig. 6. Knockdown of Ocln protein with si-RNA in the colon of control rats leads to increased visceral pain.

Fig. 6

Control rats were treated intrarectally with Ocln-specific siRNA (Ocln siRNA, n=3) or non-specific, scrambled siRNA (Ctrl siRNA, n=3). A) VMR to CRD was measured one day before treatment and 48 hrs after treatment. Each rat served as its own control, with VMR expressed as the percent of the pre-treatment 60 mmHg distention value. After VMR to CRD measurements, colon epithelium was collected from the distal (D) and proximal (P) colon for each rat. Epithelial extracts were analyzed in duplicate by Western blot at two different exposures within the linear range. C and D) Western blots of Occludin (Ocln) and β-actin are shown. Blots were quantitated, using β-actin as a loading control, with ImageJ. Protein levels are expressed as a percentage of proximal colon epithelial levels. B) In a second group of rats, 48 hrs after Ocln siRNA (n=5) or control siRNA (n=5) treatment, the distal colon was removed, the serosal musculature stripped away, and the tissue put into an Ussing Chamber to determine the mucosal to serosal transport of 4 KDa FITC-dextran, apparent permeability (Papp). Some bars are too small to see. * p < 0.001, t-test with a 1% FDR.