Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Effects of pre- and postnatal protein restriction on maternal and offspring metabolism in the nonhuman primate

doi: 10.1152/ajpregu.00150.2019

Figure Lengend Snippet: TaqMan gene expression assays for quantitative PCR in NHP tissues

Article Snippet: IGFBP1 , Rh02829190_m1 , SYT7 , Hs01590513_m1.

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Gene Assay

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Effects of pre- and postnatal protein restriction on maternal and offspring metabolism in the nonhuman primate

doi: 10.1152/ajpregu.00150.2019

Figure Lengend Snippet: P210 gene expression relative to 18S

Article Snippet: IGFBP1 , Rh02829190_m1 , SYT7 , Hs01590513_m1.

Techniques: Gene Expression

Journal: JCI Insight

Article Title: Cilia proteins are biomarkers of altered flow in the vasculature

doi: 10.1172/jci.insight.151813

Figure Lengend Snippet: Flk1 mCherry Arl13b GFP double-transgenic zebrafish embryos at 29.5 hpf were subjected to 32°C temperature for 3 hours. Single cells were harvested from dechorionated embryos, and expression of cilia-specific proteins was quantified by flow cytometry in live ECs (mCherry + ) versus non-ECs (mCherry – ). Representative dot plots show the gating strategy as applied during FACS analysis to identify ECs ( A ). Stress-responsive protein Klf4 was quantified in ECs ( B ). Protein quantification was done by measuring MFI. Cilia-specific proteins were quantified in ECs ( C ) as well as non-ECs ( D ). Arl13b expression is marked by enhanced green fluorescent protein expression. For Arl13b n = 6 (for EC and non-EC); γ-tubulin n = 5 (for EC and non-EC); Ift88 n = 4 (EC) and n = 5 (non-EC); Inversin ( n = 3 for EC and non-EC); Klf4 n = 3 (for EC and non-EC). A linear mixed model was used to examine the differences between treatment group and control group within EC (mCherry + ) or non-EC (mCherry – ). Time processed nested within day was treated as random. ARL13b and Inversin expression data were log-transformed to improve fit.

Article Snippet: Primary antibodies used in this study include ARL13b (Proteintech catalog 17711-I-AP), acetylated tubulin (Sigma catalog T6793), IFT88 (Thermo Fisher Scientific catalog PA5-18467), Inversin (Proteintech catalog 10585-I-AP), Dynein (Thermo Fisher Scientific catalog MA1-070), gTubulin (GeneTex catalog GTX113286), Alk1 (Abcam catalog ab51870), KLF4 (Proteintech catalog 11880-I-AP), HO-1 (BD, catalog 566391), NRF2 (BioLegend, catalog 939202), and bactin (Sigma catalog A5441 and Cell Signaling Technology catalog 4970P).

Techniques: Transgenic Assay, Expressing, Flow Cytometry, Control, Transformation Assay

Journal: JCI Insight

Article Title: Cilia proteins are biomarkers of altered flow in the vasculature

doi: 10.1172/jci.insight.151813

Figure Lengend Snippet: HBMVECs exposed to sickle (SS) or healthy (AA) RBCs were subjected to shear stress (1 dyne/cm 2 ), and fraction of SS ( n = 6) and AA ( n = 5) RBCs adhered to ECs after stress induction was calculated, P = 0.0081 ( A ). SS and AA RBCs were tested for ARL13b cilia prior to flow and proportion of ARL13b cilia adhered to circulating SS RBCs ( n = 16) versus AA RBCs ( n = 12) were quantified, P < 0.0001 ( B ). After flow, ARL13b expression on SS RBCs ( n = 11), but not AA RBCs ( n = 6), upon interaction with ECs, P = 0.0006 ( C ). ( A – C ) Mann-Whitney-Wilcoxon test P values are provided. Representative field (magnification 63×; scale bar = 20 μm) of a smear of SS RBCs shows cilia presence on these sickle cells, detected with FITC-conjugated anti-Arl13b antibody ( D ). Western blot plot shows the detection of cilia-specific proteins in plasma samples of healthy controls (AA) versus sickle (SS). Red asterisk represents the top IFT88 band that was used for quantification ( E ). Please note that Western blots from only 4 AA and SS samples are shown in E . A separate gel for the other 6 samples was run and quantified. Quantification includes all 10 samples from each group. Cilia-specific proteins were quantified from plasma samples of healthy controls (AA) ( n = 10) versus sickle (SS) ( n = 10) and normalized against housekeeping protein bACTIN ( F ). * P < 0.05, *** P < 0.001. A 2-tailed t test or Mann-Whitney-Wilcoxon test was performed to compare between groups.

Article Snippet: Primary antibodies used in this study include ARL13b (Proteintech catalog 17711-I-AP), acetylated tubulin (Sigma catalog T6793), IFT88 (Thermo Fisher Scientific catalog PA5-18467), Inversin (Proteintech catalog 10585-I-AP), Dynein (Thermo Fisher Scientific catalog MA1-070), gTubulin (GeneTex catalog GTX113286), Alk1 (Abcam catalog ab51870), KLF4 (Proteintech catalog 11880-I-AP), HO-1 (BD, catalog 566391), NRF2 (BioLegend, catalog 939202), and bactin (Sigma catalog A5441 and Cell Signaling Technology catalog 4970P).

Techniques: Shear, Expressing, MANN-WHITNEY, Western Blot, Clinical Proteomics

Journal: JCI Insight

Article Title: Cilia proteins are biomarkers of altered flow in the vasculature

doi: 10.1172/jci.insight.151813

Figure Lengend Snippet: Human brain microvascular ECs were sham treated or treated with the NOX inhibitor apocynin ( A ), prior to exposure or not with sickle RBCs (SS RBCs) ( n = 6). Exposure of HBMVECs to SS RBCs increased ROS generation in ECs, which is dependent on NOX. Flow cytometry analysis shows ARL13b bound to SS RBCs ( n = 6) prior to (baseline) and after interaction with shear-stressed ECs pretreated with apocynin ( B ). * P < 0.05, and **** P < 0.0001. + a = ECs treated with apocynin. For A , 1-way ANOVA test was performed, and for B , Wilcoxon’s signed-rank test was performed.

Article Snippet: Primary antibodies used in this study include ARL13b (Proteintech catalog 17711-I-AP), acetylated tubulin (Sigma catalog T6793), IFT88 (Thermo Fisher Scientific catalog PA5-18467), Inversin (Proteintech catalog 10585-I-AP), Dynein (Thermo Fisher Scientific catalog MA1-070), gTubulin (GeneTex catalog GTX113286), Alk1 (Abcam catalog ab51870), KLF4 (Proteintech catalog 11880-I-AP), HO-1 (BD, catalog 566391), NRF2 (BioLegend, catalog 939202), and bactin (Sigma catalog A5441 and Cell Signaling Technology catalog 4970P).

Techniques: Flow Cytometry, Shear

Journal: American Journal of Human Genetics

Article Title: TRPV6 Variants Interfere with Maternal-Fetal Calcium Transport through the Placenta and Cause Transient Neonatal Hyperparathyroidism

doi: 10.1016/j.ajhg.2018.04.006

Figure Lengend Snippet: Pedigrees of the Six Families with TNHP and Sanger Sequencing Confirming the Identified Mutations in TRPV6 that Show Complete Cosegregation with Phenotype

Article Snippet: The membrane was incubated overnight at 4°C with 3% BSA in PBS with 0.1% Tween-20 (PBST) for blocking and then incubated with mouse anti-c- myc antibody (Sigma #M5546, 1/500 dilution in 3% BSA in PBST) or rabbit anti-TRPV6 antibody (Alomone #ACC-036, 1/500) for 2 hr at room temperature.

Techniques: DNA Sequencing

Journal: American Journal of Human Genetics

Article Title: TRPV6 Variants Interfere with Maternal-Fetal Calcium Transport through the Placenta and Cause Transient Neonatal Hyperparathyroidism

doi: 10.1016/j.ajhg.2018.04.006

Figure Lengend Snippet: Pre- and Postnatal Clinical Findings in Six Subjects with TRPV6 Variants

Article Snippet: The membrane was incubated overnight at 4°C with 3% BSA in PBS with 0.1% Tween-20 (PBST) for blocking and then incubated with mouse anti-c- myc antibody (Sigma #M5546, 1/500 dilution in 3% BSA in PBST) or rabbit anti-TRPV6 antibody (Alomone #ACC-036, 1/500) for 2 hr at room temperature.

Techniques:

Journal: American Journal of Human Genetics

Article Title: TRPV6 Variants Interfere with Maternal-Fetal Calcium Transport through the Placenta and Cause Transient Neonatal Hyperparathyroidism

doi: 10.1016/j.ajhg.2018.04.006

Figure Lengend Snippet: Functional Analysis of TRPV6 Variants

Article Snippet: The membrane was incubated overnight at 4°C with 3% BSA in PBS with 0.1% Tween-20 (PBST) for blocking and then incubated with mouse anti-c- myc antibody (Sigma #M5546, 1/500 dilution in 3% BSA in PBST) or rabbit anti-TRPV6 antibody (Alomone #ACC-036, 1/500) for 2 hr at room temperature.

Techniques: Functional Assay, Analysis, Patch Clamp, Western Blot

Journal: Frontiers in Endocrinology

Article Title: Short-Term Diet Induced Changes in the Central and Circulating IGF Systems Are Sex Specific

doi: 10.3389/fendo.2020.00513

Figure Lengend Snippet: List of Taqman probes used for RT-PCR.

Article Snippet: Insulin-like growth factor-binding protein 1 ( Igfbp1 ) , Rn00565713_m1.

Techniques: Clinical Proteomics

Journal: Frontiers in Endocrinology

Article Title: Short-Term Diet Induced Changes in the Central and Circulating IGF Systems Are Sex Specific

doi: 10.3389/fendo.2020.00513

Figure Lengend Snippet: Relative gene expression of members of the IGF system in the hypothalamus.

Article Snippet: Insulin-like growth factor-binding protein 1 ( Igfbp1 ) , Rn00565713_m1.

Techniques: Gene Expression

Journal: medRxiv

Article Title: The miR-362-3p/ BCLAF1 axis regulates cisplatin sensitivity and metastatic progression in triple-negative breast cancer

doi: 10.64898/2026.03.09.26347941

Figure Lengend Snippet: A. Negative correlation between miR-362-3p and BCLAF1 expression in TCGA breast cancer cohort. B-D. qRT-PCR analysis of BCLAF1 and SLC44A1 mRNA levels following miR-362-3p modulation in vitro . MiR-362-3p levels inversely suppressed BCLAF1 mRNA, while SLC44A1 remains unaffected. E. Western blotting demonstrating reciprocal changes in BCLAF1 protein expression, but not SLC44A1, upon miR-362-3p overexpression (OE) or inhibition. F-G. Validation of miR-362-3p-mediated regulation of BCLAF1 mRNA ( F ) and protein ( G ) in endpoint xenograft tumors. H The densitometric quantification of the 145 kDa BCLAF1 isoform (upper band). Data represent mean±SD. * and ** represent p <0.05 and p <0.01, respectively. Non-targeting controls, NTC; empty vector, EV.

Article Snippet: Membranes were incubated with the following primary antibodies: GAPDH (14C10) rabbit monoclonal antibody (Cell Signaling Technology, #2118, RRID:AB_561053), BCLAF1 polyclonal antibody (Proteintech Group, Inc, Rosemont, IL, #26809-1-AP, RRID:AB_2880642), phospho-Histone H2A.X (Ser139) (20E3) rabbit monoclonal antibody (Cell Signaling Technology, #9718, RRID:AB_2118009), and SLC44A1 polyclonal antibody (Proteintech Group, Inc, #33135-1-AP, RRID: AB_10696180).

Techniques: Expressing, Quantitative RT-PCR, In Vitro, Western Blot, Over Expression, Inhibition, Biomarker Discovery, Plasmid Preparation

Journal: Endocrinology

Article Title: Mechanisms for hypercalciuria in pseudohypoaldosteronism type II-causing WNK4 knock-in mice.

doi: 10.1210/en.2009-0951

Figure Lengend Snippet: FIG. 1. Expression of TRPV5, TRPV6, and CBP-D28k in the Wnk4D561A/ mice kidney. A, Q-PCR analysis of TRPV5 and TRPV6. Gene expression of TRPV5 and TRPV6 in Wnk4/ (white bars) and Wnk4 D561A/ (black bars) mice kidneys (n 6/group) are presented as percentage of control (control, 100%). B, Specificity of the anti-TRPV5 antibody. Because TRPV5 expressed in MDCK cells was tagged with EGFP (32kD; right panel), the molecular weight of native TRPV5 in the mice kidney (dashed arrow) is smaller than that of mice EGFP-TRPV5 expressed in MDCK cells (solid arrow) (left panel). Anti-EGFP antibody clearly identifies the EGFP-TRPV5 protein expressed in MDCK cells but not the native TRPV5 present in mice kidney (right panel). C, IB of TRPV5, TRPV6, and CBP-D28k. IB of TRPV5 (upper panel), TRPV6 (middle panel) in crude membrane fractions, and CBP-D28k (lower panel) in whole homogenates without the nuclear fraction from Wnk4/ and Wnk4D561A/ mice kidneys (n 5/group). Semiquantification by densitometry analysis is shown. *, P 0.05 in Wnk4D561A/ vs. Wnk4/ comparison.

Article Snippet: The primary antibodies used for IB and IF were as follows: antiTRPV5 (1:200) (for IB, Santa Cruz Biotechnology, Inc., Santa Cruz, CA; for IF, Alpha Diagnostic Intl., Inc., San Antonio, TX), TRPV6 (1:200) (for IB, Alomone Labs Ltd., Jerusalem, Israel; for IF, Alpha Diagnostic Intl., Inc.), Na -K -2Cl cotransporter 2 (NKCC2) (1:200) Alpha Diagnostic Intl., Inc.), Na /H exchanger 3 (NHE3) (1:200) (Alpha Diagnostic Intl., Inc.), CBPD28k (1:10,000) (Swant, Bellinzona, Switzerland), NCC (1:200) (Chemicon, Billerica, MA), epithelium Na channel subunit [ENaC( )] (1:200) (Alomone Labs Ltd.), and aquaporin 2 (1: 200) (Santa Cruz Biotechnology, Inc.).

Techniques: Expressing, Gene Expression, Control, Molecular Weight, Membrane, Comparison

Journal: Endocrinology

Article Title: Mechanisms for hypercalciuria in pseudohypoaldosteronism type II-causing WNK4 knock-in mice.

doi: 10.1210/en.2009-0951

Figure Lengend Snippet: FIG. 2. IF staining for TRPV5, TRPV6, and CBP-D28k. A, Representative IF micrographs of TRPV5 (green; upper panels) merged with CBP-D28k (red; lower panels) in the cortex of Wnk4/ and Wnk4D561A/

Article Snippet: The primary antibodies used for IB and IF were as follows: antiTRPV5 (1:200) (for IB, Santa Cruz Biotechnology, Inc., Santa Cruz, CA; for IF, Alpha Diagnostic Intl., Inc., San Antonio, TX), TRPV6 (1:200) (for IB, Alomone Labs Ltd., Jerusalem, Israel; for IF, Alpha Diagnostic Intl., Inc.), Na -K -2Cl cotransporter 2 (NKCC2) (1:200) Alpha Diagnostic Intl., Inc.), Na /H exchanger 3 (NHE3) (1:200) (Alpha Diagnostic Intl., Inc.), CBPD28k (1:10,000) (Swant, Bellinzona, Switzerland), NCC (1:200) (Chemicon, Billerica, MA), epithelium Na channel subunit [ENaC( )] (1:200) (Alomone Labs Ltd.), and aquaporin 2 (1: 200) (Santa Cruz Biotechnology, Inc.).

Techniques: Staining