Quantification of TRPV4, AQP5, and Myosin Isoforms

Scramble control, shTRPV4 or shAQP5 cells were grown to 95% confluency and their total RNA was isolated using Direct-zol RNA isolation kit (Zymo Research) according to manufacturer’s recommendations. Reverse transcription and qPCR were performed using standard techniques described previously^{19 (link)} using the following primer sets: TRPV4: F, 5′-CCCGTGAGAACACCAAGTTT-3′ and R, 5′-GTGTCCTCATCCGTCACCTC-3′; AQP5: F, 5′-CAGCTGGCACTCTGCATCTT-3′ and R, 5′-TGAACCGATTCATGACCACC-3′; MIIA: F, 5′-ATCCTGGAGGACCAGAACTGCA-3′ and R, 5′-GGCGAGGCTCTTAGATTTCTCC-3′; MIIB: F, 5′-GCTGATGGCAACTCTCCGAAAC-3′) and R, 5′-CTTCCAGGACACCATTACAGCG-3′); MIIC: F, 5′-CAGCCGTCAAATGCAAACCGAG-3′ and R, 5′-TTGCCTCTGTCGTCACCTTCTC; 18S: F, 5′-CAGCCACCCGAGATTGAGCA-3′ and R, 5′-TAGTAGCGACGGGCGGTGTG-3′.