Figure 7. Cl^- currents in the non-ciliated cells of the efferent ductules through CFTR.

(A, D and F) Corresponding I-V curves of the whole-cell Cl^- I_ADGRG2-ED currents recorded in Figure 6 and (A, D and F) Corresponding I-V curves of the whole-cell Cl- IADGRG2-ED currents recorded in Figure 7—figure supplement 1(A,F) and Figure 7—figure supplement 3(D). WT (n = 6), Adgrg2^-/Y (n = 6); WT +CFTRinh-172 (n = 6), Adgrg2^-/Y+CFTRinh-172 (n = 6), WT +ANI9 (n = 6), Adgrg2^-/Y+ANI9 (n = 6), WT +DIDS (n = 6), Adgrg2^-/Y+DIDS (n = 6); WT +Control RNAi (n = 6), WT +CFTR RNAi (n = 6), Adgrg2^-/Y+Control RNAi (n = 6), Adgrg2^-/Y+CFTR RNAi (n = 6); WT +FSK + IBMX (n = 6), Adgrg2^-/Y+FSK+IBMX (n = 6). (B，E and G) Corresponding bar graph depicting the average current densities (pA/pF) measured at 100 mV in (A), (D) and (F). (C) qRT-PCR analysis of CFTR levels in the efferent ductules treated with CFTR siRNA (n = 3) or control RNAi (n = 3). (B, E and G) *p<0.05, **p<0.01, ***p<0.001, Adgrg2^-/Y mice compared with WT mice. #p<0.05, ##p<0.01, ###p<0.001. Treatment with selective inhibitors, stimulators or CFTR RNAi was compared with control vehicles or control RNAi. n.s., no significant difference. At least three independent biological replicates were performed for Figure 7B,E and G.

Figure 7—figure supplement 1. Effects of different stimulators or inhibitors of osmotic drivers on I _ADGRG2-ED Cl^- currents of efferent ductule cells derived from Adgrg2^-/Y mice and their wild type littermates.

(A) The whole cell Cl^- current of I _ADGRG2-ED elicited by voltage steps between −100 mV and +100 mV in a representative ADGRG2-promoter-RFP-labeled efferent ductule cells derived from the Adgrg2^-/Y mice and their wild-type littermates with or without selective inhibitors or stimulators. (B) Corresponding bar graph of average reversal potential(E_rev) (±s.e.m., n = 6 for each condition) in (A) and calculated Nernst potential at according Cl^- concentrations. n.s., no significant difference; compared to calculated Nernst potential.

Figure 7—figure supplement 2. Effects of Cl^- concentration change and CFTRinh-172 on the I_ADGRG2-ED Cl^- currents.

(A) The whole cell Cl^- current of I_ADGRG2-ED elicited by voltage steps between −100 mV and +100 mV in a representative ADGRG2-promoter-RFP-labeled efferent ductule cells derived from the Adgrg2^-/Y mice and their wild-type littermates with or without CFTR selective inhibitors CFTRinh-172, and in response to bath Cl^- concentration change (Cl^- was substituted for gluconate (Gluc^-)). (B) Representative whole cell Cl^- current of I_ADGRG2-ED versus voltage (I–V) relationships in response to voltage ramps recorded in (A) with a CsCl pipette solution. (C) Corresponding bar graph of average current desnities (pA/pF) measured at 100 mV. **p<0.01, ***p<0.001, compared with WT mice in 148.5mM Cl^- condition. #p<0.05, WT mice in 148.5mM Cl^- condition treated with CFTRinh-172 was compared with WT mice in 48.5mM Cl^- condition treated with CFTRinh-172. (D) Corresponding bar graph of average reversal potential(E_rev) (±s.e.m., n = 8 for each condition) in (A–B).

Figure 7—figure supplement 3. Effects of CFTR knocked down on the I _ADGRG2-ED Cl^- currents.

(A) The whole cell Cl^- current of I _ADGRG2-ED elicited by voltage steps between −100 mV and +100 mV of primary efferent ductile cells after CFTR-siRNA or Scramble-siRNA treatment. (B) Corresponding bar graph of average reversal potential(E_rev) (±s.e.m., n = 6 for each condition) in (A). n.s., no significant difference; compared to calculated Nernst potential.