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. 2014 Sep;93(9):911–917. doi: 10.1177/0022034514544507

Figure 1.

Figure 1.

Expression of transient receptor potential channels in cultured human odontoblast-like cells (hOBs). (a) Reverse transcription polymerase chain reaction with primers selective for GAPDH (product size = 425 bp), TRPA1 (381 bp), TRPV1 (452 bp), and TRPV4 (512/692 bp) produced bands, but no product of TRPM8 cDNA was expanded. (b) Primers selective for alkaline phosphatase (ALP; 196 bp) and collagen 1 (COL-1; 213 bp) produced bands from hOB cDNA, whereas primers selective for TRPM8 produced a product only from CHO-TRPM8 cDNA (681 bp) but not from hOBs. (c) Lysates (30 µg of total protein) from hOBs (OB), wild-type mouse urothelium (U+), and TRPV4 –/– mouse urothelium (U–) showed expression of GAPDH (37 kDa, left blot). Lysates from hOBs and wild-type mouse urothelium showed the double band corresponding to TRPV4 (98 and 104 kDa, right blot), but it was absent in TRPV4 –/– mouse urothelium. (d) Immunostaining of hOBs with rabbit anti-TRPV4 (1:200) and donkey anti-rabbit-Alexa 546 (1:1,000) revealed positive TRPV4 immunoreactivity that was not absent in the “no primary” cells. Images are representative of data obtained from n = 3 independent experiments.