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: Analysis

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: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: Cytosolic and mitochondrial calcium concentration of TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM cisplatin. ( A ) Immunostaining of transient receptor potential ankyrin 1 (TRPA1)- or transient receptor potential vanilloid 1 (TRPV1)-positive DRG neurons and live imaging of Fluo-4 (green) and Rhod-2 (red). ( B ) The cytosolic calcium of TRPV1-positive DRG neurons concentration instantly increased to 1.067 ± 0.018 (* p < 0.05) after 5 min and continuously increased to 1.394 ± 0.068 (** p < 0.01) after 70 min. The mitochondrial calcium concentration declined to 1.095 ± 0.009 (* p < 0.05) after 5 min for TRPV1-positive DRG neurons and steadily declined to 0.577 ± 0.044 after 70 min (** p < 0.01). ( C ) During the exposure of 10 µM cisplatin, the cytosolic calcium concentration of TRPA1-positive neurons increased after 5 min to 1.024 ± 0.011 (** p < 0.01) until 70 min at 1.240 ± 0.074 (*** p < 0.001). The mitochondrial calcium concentration was lower after 10 min at 0.928 ± 0.029 (*** p < 0.001) until it reached 0.790 ± 0.050 at 70 min (*** p < 0.001). ( D ) The cytosolic calcium concentration of TRPV1-positive DRG neurons was increased after 15 min to 65 min compared to TRPA1-positive DRG neurons (* p < 0.05, ** p < 0.01). ( E ) The mitochondrial calcium concentration of TRPA1- or TRPV1-positive DRG neurons was different after 5 min until the end of the experiment (* p < 0.05, ** p < 0.01). TRPV1-positive sensory neurons had a lower mitochondrial calcium concentration during exposure to 10 µM cisplatin after 15 min (* p < 0.05, ** p < 0.01). n = 6 cells per condition. * = significant effect between TRPA1+ and TRPV1+ DRG neurons. Scale = 50 µm.

Article Snippet: The cells were first stained with extracellular TRPA1 or TRPV1 antibodies (Alomone Labs, Jerusalem, Israel).

Techniques: Concentration Assay, Immunostaining, Imaging

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: Cytosolic and mitochondrial calcium concentration of TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM oxaliplatin. ( A ) Immunostaining of TRPA1- or TRPV1-positive DRG neurons and live imaging of Fluo-4 (green) and Rhod-2 (red) before and after application of 10 µM cisplatin or oxaliplatin. ( B ) TRPA1-positive DRG neurons showed an immediate increase of the cytosolic calcium concentration to 1.037 ± 0.006 (** p < 0.01) after 5 min and continuously increased to 1.372 ± 0.062 after 70 min (*** p < 0.001). The mitochondrial calcium concentration decreased after 5 min to 0.967 ± 0.008 (*** p < 0.001) and declined to 0.712 ± 0.010 after 70 min (*** p < 0.001). ( C ) The TRPV1-positive DRG neurons showed an immediate increase of the cytosolic calcium concentration to 1.049 ± 0.005 (*** p < 0.001) after 5 min and increased to 1.480 ± 0.034 after 70 min (*** p < 0.001). The mitochondrial calcium concentration decreased to 0.959 ± 0.020 (* p < 0.05) after 5 min and to 0.725 ± 0.077 (** p < 0.01) after 70 min (* p < 0.05). ( D ) No difference could be determined in the cytosolic calcium concentration of TRPA1- or TRPV1-positive DRG neurons ( p > 0.05). ( E ) No difference could be determined in the mitochondrial calcium concentration of TRPA1- or TRPV1-positive DRG neurons ( p > 0.05). n = 6 cells per condition. * = significant effect between TRPA1+ and TRPV1+ DRG neurons. Scale = 50 µm.

Article Snippet: The cells were first stained with extracellular TRPA1 or TRPV1 antibodies (Alomone Labs, Jerusalem, Israel).

Techniques: Concentration Assay, Immunostaining, Imaging

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: Relative cytosolic and mitochondrial calcium concentration during exposure to 10 µM cis- or oxaliplatin of TRPA1- or TRPV1-positive DRG neurons. ( A ) No differences in the cytosolic calcium concentration of TRPA1-positive DRG neurons after exposure to 10 µM cis- or oxaliplatin could be determined ( p > 0.05). ( B ) The cytosolic calcium of TRPV1-positive DRG neurons concentration during exposure to cis- or oxaliplatin showed no difference ( p > 0.05). ( C ) During exposure to 10 µM cis- or oxaliplatin, no difference in the mitochondrial calcium concentration of TRPA1-positive DRG neurons could be determined ( p > 0.05). ( D ) While exposing TRPV1-positive DRG neurons, differences between the two chemotherapeutics on the mitochondrial calcium concentration could be determined after 20 min. The mitochondrial calcium concentration of TRPV1-positive DRG neurons exposed to oxaliplatin was higher at 0.856 ± 0.033 compared to 0.727 ± 0.040 (* p < 0.05). The mitochondrial calcium concentration was higher for oxaliplatin-exposed TRPV1-positive DRG neurons until 0.786 ± 0.058 after 40 min (* p < 0.05). n = 6 cells per condition. * = significant effect between cis- and oxaliplatin, ** p < 0.01

Article Snippet: The cells were first stained with extracellular TRPA1 or TRPV1 antibodies (Alomone Labs, Jerusalem, Israel).

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: ROS production of TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM cisplatin. ( A ) Fluorescence of CellRox (green) in TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM cisplatin at different time points. ( B ) TRPA1-positive sensory neurons showed an instant increase of the ROS production to 1.431 ± 0.042 (*** p < 0.001) after 5 min and reached its peak at 2.100 ± 0.197 after 30 min (*** p < 0.001). After 50 min, the ROS production dropped below the control level at 50 min to 0.607 ± 0.036 (*** p < 0.001) and further declined to 0.370 ± 0.035 (*** p < 0.001) after 70 min. ( C ) After exposure to 10 µM cisplatin of the TRPV1-positive DRG neurons, the ROS production increased to 1.238 ± 0.042 (*** p < 0.001) after 5 min and dropped below control level at 50 min at 0.873 ± 0.068 (* p < 0.05) and steadily declined to 0.756 ± 0.072 (** p < 0.01) after 70 min. n = 6 cells per condition. * = significant effect compared to control level. Scale = 50 µm.

Article Snippet: The cells were first stained with extracellular TRPA1 or TRPV1 antibodies (Alomone Labs, Jerusalem, Israel).

Techniques: Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: ROS production of TRPA1- or TRPV1-positive DRG neurons during exposure to 10 µM oxaliplatin. ( A ) Fluorescence of CellRox (green) of TRPA1- or TRPV1-positive DRG neurons at different time points. ( B ) The ROS production of TRPA1-positive DRG neurons increased after 5 min to 1.421 ± 0.103 (** p < 0.01) during exposure to 10 µM oxaliplatin and dropped to control level after 40 min at 1.31 ± 0.194 ( p > 0.05). ( C ) The TRPV1-positive sensory neurons had an increased ROS production after 5 min at 1.080 ± 0.020 (** p < 0.01) and dropped to the control level after 10 min at 1.033 ± 0.036 ( p > 0.05). After 30 min, the ROS production declined below the control level at 0.810 ± 0.079 (* p < 0.05) and steadily declined to 0.429 ± 0.050 after 70 min (*** p < 0.001). n = 6 cells per condition. * = significant effect compared to control level. Scale = 20 µm.

Article Snippet: The cells were first stained with extracellular TRPA1 or TRPV1 antibodies (Alomone Labs, Jerusalem, Israel).

Techniques: Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: The ROS production of TRPA1- or TRPV1-positive DRG neurons during exposure to cis- or oxaliplatin. ( A ) A difference could be determined for TRPA1-positive DRG neurons at 30 min with a higher ROS production for cisplatin-exposed DRG neurons at 2.100 ± 0.197 (* p < 0.05). After 45 min (1.194 ± 0.223; * p < 0.05) until 70 min (1.022 ± 0.233; * p < 0.05), the ROS production was higher for oxaliplatin-exposed neurons. ( B ) The ROS production of TRPV1-positive DRG neurons was higher for cisplatin-exposed neurons over the entire experimental period (* p < 0.05, ** p < 0.01). n = 6 cells per condition. * = significant effect compared to control level.

Article Snippet: The cells were first stained with extracellular TRPA1 or TRPV1 antibodies (Alomone Labs, Jerusalem, Israel).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Platinum-Based Drugs Cause Mitochondrial Dysfunction in Cultured Dorsal Root Ganglion Neurons

doi: 10.3390/ijms21228636

Figure Lengend Snippet: Antibodies and dilution used for the TRPA1- and TRPV1-positive staining.

Article Snippet: The cells were first stained with extracellular TRPA1 or TRPV1 antibodies (Alomone Labs, Jerusalem, Israel).

Techniques: Staining