(A) Representative Western blots illustrating PC-1, PC-2, SK3, IK, TRPV4, Piezo1, eNOS, GPR68, and actin proteins in mesenteric arteries of Pkd1fl/fl and Pkd1 ecKO mice. (B) Mean data for PC-1, …
Representative of four separate experiments.
(A) Representative traces illustrating reversible flow-mediated dilation in pressurized (80 mmHg) mesenteric arteries of Pkd1fl/fl and Pkd1 ecKO mice. (B) Mean dilation to flow (15 dyn/cm2) or ACh …
(A) Representative traces illustrating dilations to ACh (10 µM) and Ca2+-free bath solution in pressurized (80 mmHg) Pkd1fl/fl and Pkd1 ecKO arteries. (B) PC-1 knockout attenuates flow-mediated …
(A) Blood pressure recordings obtained over 24 hr in a Pkd1fl/fl and Pkd1 ecKO mouse. (B) Mean arterial pressures (MAPs) in Pkd1fl/fl (n=5) and Pkd1 ecKO (n=7) mice. *p<0.05 versus Pkd1fl/fl. (C) …
(A) Mean diastolic and systolic blood pressures for Pkd1fl/fl (n=5) and Pkd1 ecKO (n=7) mice. *=p<0.05 versus Pkd1fl/fl. (B) Mean data for Pkd1fl/fl (n=5) and Pkd1 ecKO (n=7) mouse activity.
(A) Representative Western blots illustrating the immunoblot (IB) detection of both PC-1 and PC-2 in PC-2 immunoprecipitate (IP) (n=5). (B) PC-1 (Alexa546) and PC-2 (Alexa488) antibody labeling …
PC-1 (Alexa546) and PC-2 (Alexa488) antibody labeling and FRET in mesenteric artery ECs of Pkd1 ecKO (A) and Pkd2 ecKO (B) mice. Scale bars=10 µm. immunoFRET, immunofluorescence energy transfer.
(A) TIRF-SMLM images of PC-1 and PC-2 surface clusters in a Pkd1fl/fl and Pkd1 ecKO EC. Scale bars=5 µm. (B) Mean data for PC-1 and PC-2 cluster density measured in Pkd1fl/fl and Pkd1 ecKO ECs under …
Mean data illustrating the localization precision (FWHM) of Alexa Fluor 488 (A)- and Alexa Fluor 647 (B)-tagged secondary antibodies in ECs under all conditions when imaged using SMLM. (A) …
(A) Histogram illustrating PC-1 cluster size distribution in a Pkd1fl/fl (static) and Pkd1 ecKO (static) EC. (B) Histogram illustrating PC-2 cluster sizes in a Pkd1fl/fl (static) and Pkd1 ecKO …
(A) TIRF-SMLM images of PC-1 and PC-2 surface clusters in a Pkd2fl/fl and Pkd2 ecKO EC. Scale bars = 5 µm. (B) Mean data for PC-1 and PC-2 cluster density measured in Pkd2fl/fl and Pkd2 ecKO ECs …
(A) TIRF-SMLM images of PC-1 and PC-2 clusters, overlap of PC-1 and PC-2 data and overlap of PC-1 and PC-2 data following Costes’ randomization simulation. Scale bars = 5 µm. (B) Mean experimental …
(A) Representative Western blots of PC-1, PC-2, and eNOS in mesenteric arteries of Pkd1fl/fl/Pkd2fl/fl and Pkd1/Pkd2 ecKO mice. (B) Mean data. *p<0.05 versus Pkd1fl/fl/Pkd2fl/fl. n=5 for each data …
(A) Representative traces illustrating 60 mM-induced K+ constriction in a pressurized (10 mmHg) Pkd1fl/fl/Pkd2fl/fl and Pkd1/Pkd2 ecKO artery. (B) Mean data for 60 mM K+-induced constriction. n=6 …
Original recordings illustrating that intracellular introduction via the patch pipette of either a PC-1 scrambled (scram) peptide or a PC-2 scrambled peptide does not alter flow (10 …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Strain, strain background (Mus musculus) | Pkd1fl/fl | Baltimore PKD Core Center | PMID:15579506MGI: 3617325 | Mice with Pkd1 gene flanked by loxP regions. |
Strain, strain background (M. musculus) | Pkd2fl/fl | Baltimore PKD Core Center | PMID:20862291MGI: 4843127 | Mice with Pkd2 gene flanked by loxP regions. |
Strain, strain background (M. musculus) | Pkd1fl/fl /Pkd2fl/fl | This paper | Mouse line created in-house by mating Pkd1fl/fl with Pkd2fl/fl. Mice with Pkd1 and Pkd2 genes flanked by loxP regions. | |
Strain, strain background (M. musculus) | Cdh5(PAC)-CreERT2 | Cancer Research UK | RRID: MGI: 3848982 | Mice with tamoxifen-inducible Cre recombinase that is expressed specifically in endothelial cells. |
Strain, strain background (M. musculus) | Pkd1fl/fl: Cdh5(PAC)-CreERT2 | This paper | Mouse line created in-house by mating Pkd1fl/fl with Cdh5(PAC)-CreERT2. Mice with inducible endothelial cell-specific deletion of PC-1. | |
Strain, strain background (M. musculus) | Pkd2fl/fl: Cdh5(PAC)-CreERT2 | PMID:32364494 | Mouse line created in-house by mating Pkd2fl/fl with Cdh5(PAC)-CreERT2. Mice with inducible endothelial cell-specific deletion of PC-2. | |
Strain, strain background (M. musculus) | Pkd1fl/fl/Pkd2fl/fl: Cdh5(PAC)-CreERT2 | This paper | Mouse line created in-house by mating Pkd1fl/fl Cdh5(PAC)-CreERT2 with Pkd2fl/fl Cdh5(PAC)-CreERT2. Mice with inducible endothelial cell-specific deletion both PC-1 and PC-2. | |
Antibody | Anti-PC-1 (mouse monoclonal) | Baltimore PKD Core | Mouse mAB SF4AZ E3 | IF (1:100)Lattice SIM (1:100)SMLM (1:100) |
Antibody | Anti-PC-2 (rabbit monoclonal) | Baltimore PKD Core | Rabbit mAB 3374 CT-14/4 | IF (1:100)Lattice SIM (1:100)SMLM (1:100) |
Antibody | Anti-CD31 (rat monoclonal) | Abcam | Cat. #: Ab7388RRID: AB_305905 | IF (1:100)Lattice SIM (1:100)SMLM (1:100) |
Antibody | Anti-PC-1 (H-260)(rabbit polyclonal) | Santa Cruz Biotechnology | Cat. #: sc-25570, RRID: AB_2163357 | N-FRET (1:100) |
Antibody | Anti-PC-2 (mouse monoclonal) | Santa Cruz Biotechnology | Cat. #: sc-28331, RRID: AB_672377 | N-FRET (1:100) |
Antibody | Alexa 488-conjugated anti-rat IgG (donkey polyclonal) | Thermo Fisher Scientific | Cat. #: A-21208,RRID: AB_2535794 | IF (1:500) |
Antibody | Alexa 546-conjugated anti-mouse IgG (donkey polyclonal) | Thermo Fisher Scientific | Cat. #: A-10036,RRID: AB_2534012 | IF (1:500) |
Antibody | Alexa 488-conjugated anti-mouse IgG (donkey polyclonal) | Thermo Fisher Scientific | Cat. #: A-21202,RRID: AB_141607 | Lattice SIM (1:500) SMLM (1:500) |
Antibody | Alexa 546-conjugated anti-rabbit IgG (donkey polyclonal) | Thermo Fisher Scientific | Cat. #: A-10040,RRID: AB_2534016 | Lattice SIM(1:500) |
Antibody | Alexa 647-conjugated anti-rat IgG (goat polyclonal) | Thermo Fisher Scientific | Cat. #: A-21247,RRID: AB_141778 | Lattice SIM(1:500) |
Antibody | Alexa 555-conjugated anti-rat IgG (goat polyclonal) | Thermo Fisher Scientific | Cat. #: A-21434,RRID: AB_2535855 | SMLM (1:500) |
Antibody | Alexa 647-conjugated anti-rabbit IgG (donkey polyclonal) | Thermo Fisher Scientific | Cat. #: A-31573,RRID: AB_2536183 | SMLM (1:500) |
Antibody | Alexa Fluor 488-conjugated anti-mouse (goat polyclonal) | Thermo Fisher Scientific | Cat. #: A-11001, RRID: AB_2534069 | N-FRET (1:100) |
Antibody | Alexa Fluor 555-conjugated anti-rabbit (donkey polyclonal) | Thermo Fisher Scientific | Cat. #: A-31572, RRID: AB_162543 | N-FRET (1:100) |
Antibody | Anti-PC-1 (mouse monoclonal) | Santa Cruz Biotechnology | Cat. #: sc-130554,RRID: AB_2163355 | WB (1:100) |
Antibody | Anti-PC-2 (mouse monoclonal) | Santa Cruz Biotechnology | Cat. #: sc-47734,RRID: AB_672380 | WB (1:100)IP (1:20) |
Antibody | Anti-IK1 (D-5) (mouse monoclonal) | Santa Cruz Biotechnology | Cat. #: sc-365265,RRID: AB_10841432 | WB (1:100) |
Antibody | Anti-SK3(rabbit polyclonal) | Abcam | Cat. #: ab28631,RRID: AB_775888 | WB (1:100) |
Antibody | Anti-eNOS [M221](mouse monoclonal) | Abcam | Cat. #: ab76198,RRID: AB_1310183 | WB (1:100) |
Antibody | Anti-TRPV4(clone 1B2.6) (mouse monoclonal) | Millipore Sigma | Cat. #: MABS466 | WB (1:100) |
Antibody | Anti-Actin (clone C4)(mouse monoclonal) | MilliporeSigma | Cat. #: MAB1501,RRID: AB_2223041 | WB (1:1000) |
Antibody | Anti-p-eNOS (rabbit polyclonal) | Cell Signaling Technology | Cat. #: 9571,RRID: AB_329837 | WB (1:100) |
Antibody | Anti-GPR68(rabbit polyclonal) | NOVUS Biologicals | Cat. #: NBP2-32747 | WB (1:100) |
Antibody | Anti-Piezo1 (rabbit polyclonal) | ProteinTech | Cat. #: 15939-1-AP,RRID: AB_2231460 | WB (1:100) |
Antibody | Mouse IgG control (mouse polyclonal) | MilliporeSigma | Cat. #: 12-371,RRID: AB_145840 | IP (1:20) |
Peptide, recombinant protein | PC-1 coiled-coil domain peptide | Genscript | FDRLNQATE DVYQLEQQL | 1 µM |
Peptide, recombinant protein | PC-1 scrambled peptide | Genscript | QLNDLFQTE VAEDLQRYQ | 1 µM |
Peptide, recombinant protein | PC-2 coiled-coil domain peptide | Genscript | KRREVLGRLL | 1 µM |
Peptide, recombinant protein | PC-2 scrambled peptide | Genscript | VLKLLRRRGE | 1 µM |
Commercial assay or kit | Pierce crosslink magnetic IP/co-IP Kit | Thermo Fisher Scientific | Cat. #: 88805 | |
Other | DAPI stain | Thermo FisherScientific | Cat. #: 3571RRID: AB_2307445 | 1:1000 |
Unlabeled source data.
Labeled source data.