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. 2019 May 7;33(8):9182–9193. doi: 10.1096/fj.201900321R

Figure 6.

Figure 6

Sialic acid–recognizing CBM can inhibit TRPC6 like sKlotho. A) Production and purification of CBM. His-tagged CBM was subcloned in pEHISTEVb vector, expressed in bacteria, and purified with a Ni–nitrilotriacetic acid column. Coomassie blue gel of proteins in whole-cell lysate (WCL), supernatant of lysate after centrifugation (S), flow-through of after loading on to the column (FT), and fractions 4, 25, 29, 32, 36, 40. MW markers are in kilodaltons. B) Incubation with CBM resulted in inhibition of TRPC6 channel currents expressed in HEK cells dose dependently. The effect of sKlotho (sKL; 1 nM) is shown for comparison. Bars are means ± sem of inward current density at −100 mV. Inset shows I-V relationship curves. Veh, vehicle. C) Incubation with CBM-inhibited TRPC6 in FGFR-null L6 cells. D) No additivity between sKL (1 nM) and CBM (1 nM) on TRPC6 currents in L6 cells. n = 7–10 each group. *P < 0.05 vs. control (first bar from left, no sKL, no CBM).