Research Paper Volume 12, Issue 12 pp 11485—11499

Figure 6. Overexpression of CTRP13 elevated the activation of CaMKKβ/AMPK pathway and inhibited the expression of LN and CAV-1 in rLSECs. Cells were transfected with LV-CTRP13 or LV-CON and blocked with CaMKK or AMPK inhibitor. All samples were treated with 25 mg/ml high glucose for 24 h. Untreated intact samples were run as a control in each experiment. (A) qRT-PCR analysis of CTRP13 mRNA. (B) qRT-PCR analysis of CaMKKβ mRNA. (C) qRT-PCR analysis of AMPKα mRNA. (D) qRT-PCR analysis of LN mRNA. (E) qRT-PCR analysis of CAV-1 mRNA. The results were normalised to GAPDH mRNA levels. (F) The protein expression levels of each group were detected using western blotting, and GAPDH was used as a loading control. (G) Western blotting results showing relative CTRP13 expression. (H) Western blotting results showing relative CaMKKβ expression. (I) Western blotting results showing relative p-CaMKKβ expression of CaMKKβ activation. (J) Western blotting results showing relative AMPKα expression. (K) Western blotting results showing relative p-AMPKα expression of AMPKα activation. (L) Western blotting results showing relative LN expression. (M) Western blotting results showing relative CAV-1 expression. GAPDH (37 kDa) was used as a loading control. All results are expressed as mean±S.D. from three independent experiments, ^*P < 0.05, ^**P < 0.01, ^***P < 0.001vs. control. ^#P < 0.05, ^###P < 0.001 vs the high glucose + LV-CON group. ^χP < 0.05, ^χχP < 0.01, ^χχχP < 0.001 vs the high glucose + LV-CTRP13 group, respectively.