Figure 2. Lnc-MGC and cluster miRNAs are upregulated under diabetic conditions.

(a) Primers for almost all (∼40) miRNAs in miR-379 cluster were designed and the expression of 40 miRNAs were confirmed by RT-PCRs and found to be increased significantly, Benjamini–Hochberg adjusted P<0.05 (ref. ⁶⁹) in glomeruli from diabetic mice (STZ, STZ-injected type 1 model, blue bars; db/db, type 2 diabetic model, red bars) compared with respective non-diabetic mice (vehicle injected or db/+). Five mice in each group. Results are mean+s.e. from triplicate PCRs. No significant change was detected in the expression of miR-882 (outside of the miR-379 cluster). (b) 5′RACE of the region upstream of miR-379 identified the typical consensus sequence of initiator (INR; PyPyANT/APyPy) at the transcription start site. Smad-binding elements (CAGA repeats) and CHOP binding consensus and E-box sequences were found upstream of miR-379. (c) The expression of lnc-MGC in glomeruli from diabetic mice (STZ and db/db) compared with corresponding controls (vehicle injected CTR and db/+). Five mice in each group. *P<0.05, (d) The expression of lnc-MGC and cluster miRNAs in MMC treated with 25 mM HG (72 h) or 10 ng ml⁻¹ TGF-β1 (24 h) compared with respective controls normal glucose (5.5 mM) or SD. respectively. (e) Significant lower expression of potential targets of miR-379 cluster in kidney glomeruli from diabetic mice (type 2 db/db and type 1 STZ) than the respective non-diabetic control mice. Five mice in each group. *P<0.05. (f) Decrease of potential targets of miR-379 cluster in MMC treated with TGF-β1 or HG. For experiments with MMC, results are mean+s.e. in triplicate PCRs from three independent culture experiments. *P<0.05.