Lmd system lmd6000

Laser‐capture microdissection (LMD) was performed using a Leica LMD system (LMD6000), as described previously.18 Briefly, formalin‐fixed paraffin‐embedded tissues were cut into 10‐μm‐thick sections, mounted on polyethylene terepthalate membrane slides, and stained with hematoxylin–eosin. Next, proximal or distal tubules in the renal cortex were microdissected using an LMD6000 microscope. The renal proximal tubule cells were identified via their unique brush border membranes.21 The other tubule cells in the renal cortex were recognized as distal tubule cells (Figure 1B). In total, 60 areas (≈500 000 μm²) of proximal or distal tubules were microdissected from the renal cortex per mouse in each group (6 mice in each group). Total RNA was extracted from microdissected tissue using the RNeasy FFPE Kit (Qiagen). The cDNA was synthesized using the SuperScript III First‐Strand System (Invitrogen) and applied to Taqman reverse transcription quantitative polymerase chain reaction analysis.