Journal:

Article Title: PKHD1, the Polycystic Kidney and Hepatic Disease 1 Gene, Encodes a Novel Large Protein Containing Multiple Immunoglobulin-Like Plexin-Transcription-Factor Domains and Parallel Beta-Helix 1 Repeats

doi:

Figure Lengend Snippet: Structure of full-length PKHD1 and its splicing variants. A, Set of 71 nonoverlapping exons that spans the entire length of PKHD1 (upper row) and 15 additional overlapping exons that use different splice sites (gray boxes, lower row). Exons, which are not present in the cDNA that encodes the longest ORF, are indicated by hatched boxes. The position of important protein domains is indicated. B, Approximate location of each primer set used to amplify various cDNAs, with representative set of amplified products (below each schema). White boxes indicate noncoding exons in the corresponding transcripts while gray boxes identify exons with alternative boundaries (A). The templates used for each amplification are as follows: human adult kidney double-stranded cDNA for primer sets 1–4, 6, and 8; human kidney mRNA and total RNA for primer sets 5 and 7; human adult kidney cDNA library for primer set 9. “SC” indicates approximate location of stop codons, and “ORF” indicates that an open reading frame extends throughout the length of the fragment. C, Longest ORF identified by RT-PCR/cDNA amplification. This ORF is the composite sequence of products 2.1 and 4.1 of panel B and includes a total of 67 exons.

Article Snippet: A second set of products were generated by RT-PCR using either 20 ng of human adult kidney mRNA (Clontech) or 1.5–4.0 μg of human adult kidney total RNA as template.

Techniques: Amplification, cDNA Library Assay, Reverse Transcription Polymerase Chain Reaction, Sequencing

Journal: Drug Design, Development and Therapy

Article Title: Astragaloside IV Promotes Osteogenic Differentiation of Periodontal Ligament Stem Cells via Activating PI3K/AKT/eNOS/NO Signaling Pathway: In vitro and in vivo Study

doi: 10.2147/DDDT.S514682

Figure Lengend Snippet: Effect of AS-IV on proliferation and osteogenic differentiation of PDLSCs. ( A ) CCK-8 analysis of AS-IV effects on PDLSCs on day 1, 3, and 5. ( B and C ) ALP activity ( B ) and extracellular matrix mineralization ( C ) quantification of PDLSCs affected by AS-IV. ( D ) ALP staining. Scale bar: 500 μm. ( E ) Alizarin Red staining. Scale bar: 500 μm. ( F – H ) qRT-PCR analysis of COL-1 (F), ALP(G), and RUNX-2 ( H ) in PDLSCs cultured with AS-IV. ( I – L ) Western blot and semi-quantitative analysis of COL-1 (J), ALP(K), and RUNX-2 ( L ) in PDLSCs cultured with AS-IV. 20 μM AS-IV group had a significant promoting effect. Data are presented as mean ± SD. ** P <0.01, *** P <0.001, **** P <0.0001. Each experiment was repeated five times.

Article Snippet: Incubate the membrane with primary antibodies toward target proteins including ALP (Proteintech, 11187-1-AP, 1:1000), RUNX-2 (ImmunoWay, YM8347, 1:2000), COL-1 (Proteintech, 67288-1-Ig, 1:5000), eNOS (Abways, AB3537, 1:500), iNOS (Abways, CY3425, 1:500), p-AKT (Abways, CY6569, 1:1000), AKT (Abways, CY5561, 1:1000) and GAPDH (Proteintech, 10494-1-AP, 1:5000) overnight at 4°C.

Techniques: CCK-8 Assay, Activity Assay, Staining, Quantitative RT-PCR, Cell Culture, Western Blot

Journal: Drug Design, Development and Therapy

Article Title: Astragaloside IV Promotes Osteogenic Differentiation of Periodontal Ligament Stem Cells via Activating PI3K/AKT/eNOS/NO Signaling Pathway: In vitro and in vivo Study

doi: 10.2147/DDDT.S514682

Figure Lengend Snippet: Effect of LY294002 on osteogenic differentiation and pathway-associated factor of PDLSCs following AS-IV incubation. ( A and D ) ALP staining( A ) and ALP activity quantification( D ) of PDLSCs affected by LY294002. Scale bar: 500 μm. ( B and E ) Immunofluorescence staining( B ) of COL-1 (green) and DAPI staining (blue) and quantification(E). Scale bar: 50 μm. ( C and F ) Immunofluorescence staining( C ) of eNOS (green) and DAPI staining (blue) and quantification(F). Scale bar: 50 μm. ( G ) NO2- concentration represented the level of NO in cell supernatants. ( H–K ) Western blot and semi-quantitative analysis of COL-1(I), ALP(J), and RUNX-2 ( K ) in PDLSCs cultured with AS-IV and LY294002. ( L–O ) Western blot and semi-quantitative analysis of eNOS(M), iNOS( N ) and p-AKT/AKT ( O ) after addition of AS-IV at different time points. ( P–S ) Western blot and semi-quantitative analysis of eNOS(Q), iNOS( R ) and p-AKT/AKT ( S ) in PDLSCs cultured with AS-IV and LY294002. Data are presented as mean ± SD. * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001. Each experiment was repeated five times.

Article Snippet: Incubate the membrane with primary antibodies toward target proteins including ALP (Proteintech, 11187-1-AP, 1:1000), RUNX-2 (ImmunoWay, YM8347, 1:2000), COL-1 (Proteintech, 67288-1-Ig, 1:5000), eNOS (Abways, AB3537, 1:500), iNOS (Abways, CY3425, 1:500), p-AKT (Abways, CY6569, 1:1000), AKT (Abways, CY5561, 1:1000) and GAPDH (Proteintech, 10494-1-AP, 1:5000) overnight at 4°C.

Techniques: Incubation, Staining, Activity Assay, Immunofluorescence, Concentration Assay, Western Blot, Cell Culture

Journal: Drug Design, Development and Therapy

Article Title: Astragaloside IV Promotes Osteogenic Differentiation of Periodontal Ligament Stem Cells via Activating PI3K/AKT/eNOS/NO Signaling Pathway: In vitro and in vivo Study

doi: 10.2147/DDDT.S514682

Figure Lengend Snippet: Effect of L-NAME on osteogenic differentiation and pathway-associated factor of PDLSCs following AS-IV incubation. ( A and B ) ALP staining( A ) and ALP activity quantification( B ) of PDLSCs affected by L-NAME. Scale bar: 500 μm. ( C and D ) Immunofluorescence staining( C ) of COL-1 (green) and DAPI staining (blue) and quantification(D). Scale bar: 50 μm. ( E–H ) Western blot and semi-quantitative analysis of COL-1 (F), ALP(G), and RUNX-2 ( H ) in PDLSCs cultured with AS-IV and L-NAME. ( I–K ) Western blot and semi-quantitative analysis of eNOS( J ) and p-AKT/AKT ( K ) in PDLSCs cultured with AS-IV and L-NAME. Data are presented as mean ± SD. * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001. ns: P> 0.05. Each experiment was repeated five times.

Article Snippet: Incubate the membrane with primary antibodies toward target proteins including ALP (Proteintech, 11187-1-AP, 1:1000), RUNX-2 (ImmunoWay, YM8347, 1:2000), COL-1 (Proteintech, 67288-1-Ig, 1:5000), eNOS (Abways, AB3537, 1:500), iNOS (Abways, CY3425, 1:500), p-AKT (Abways, CY6569, 1:1000), AKT (Abways, CY5561, 1:1000) and GAPDH (Proteintech, 10494-1-AP, 1:5000) overnight at 4°C.

Techniques: Incubation, Staining, Activity Assay, Immunofluorescence, Western Blot, Cell Culture

Journal: Drug Design, Development and Therapy

Article Title: Astragaloside IV Promotes Osteogenic Differentiation of Periodontal Ligament Stem Cells via Activating PI3K/AKT/eNOS/NO Signaling Pathway: In vitro and in vivo Study

doi: 10.2147/DDDT.S514682

Figure Lengend Snippet: Hematoxylin and eosin staining and immunohistochemical staining. ( A ) HE staining of the tension-side periodontal tissues of the control group (A-a) and AS-IV group (A-b) after OTM for 14 days. Scale bar: 50 μm. ( B ) Expression of ALP of the control group (B-a) and AS-IV group (B-b) after OTM for 14 days and quantitative analysis (B-c). Scale bar: 20 μm. ( C ) Expression of COL-1 of the control group (C-a) and AS-IV group (C-b) after OTM for 14 days and quantitative analysis (C-c). Scale bar: 20 μm. ( D ) Expression of eNOS of the control group (D-a) and AS-IV group (D-b) after OTM for 14 days and quantitative analysis (D-c). Scale bar: 20 μm. Data are presented as mean ± SD (n=8 rat/group). * P <0.05, ** P <0.01.

Article Snippet: Incubate the membrane with primary antibodies toward target proteins including ALP (Proteintech, 11187-1-AP, 1:1000), RUNX-2 (ImmunoWay, YM8347, 1:2000), COL-1 (Proteintech, 67288-1-Ig, 1:5000), eNOS (Abways, AB3537, 1:500), iNOS (Abways, CY3425, 1:500), p-AKT (Abways, CY6569, 1:1000), AKT (Abways, CY5561, 1:1000) and GAPDH (Proteintech, 10494-1-AP, 1:5000) overnight at 4°C.

Techniques: Staining, Immunohistochemical staining, Control, Expressing

Journal: Oncology letters

Article Title: Cadherin-16 inhibits thyroid carcinoma cell proliferation and invasion.

doi: 10.3892/ol.2022.13265

Figure Lengend Snippet: Figure 1. CDH16 is downregulated in TC from three GEO datasets and there is no significant variation in copy number in TCGA dataset. The fold‑change, P‑value and underexpression gene ranks are based on Oncomine 4.5 analysis. Box plots showing CDH16 mRNA levels in patients with PTC in (A) He, (B) Vasko and (C) Giordano thyroid datasets from GEO. Box plot showing CDH16 mRNA levels in patients with (D) FTC and (E) ATC in the Giordano thyroid dataset. (F) Box plot showing CDH16 copy number in TCGA thyroid dataset. N, normal; CDH, cadherin; TC, thyroid carcinoma; GEO, Gene Expression Omnibus; TCGA, The Cancer Genome Atlas; PTC, papillary thyroid cancer; FTC, follicular thyroid cancer; ATC, anaplastic thyroid cancer.

Article Snippet: The membranes were incubated overnight at 4 ̊C with primary antibodies against CDH16 (1:1,000, cat. no. 15107‐1‐AP, ProteinTech Group, Inc.), DNA polymerase (POL)D1 (1:1,000, cat. no. 15646‐1‐AP, ProteinTech Group, Inc), minichromosome maintenance (MCM)6 (1:2,000, cat. no. 13347‐2‐AP; ProteinTech Group, Inc), claudin (CLDN)1 (1:1,500, cat. no. 13050‐1‐AP, ProteinTech Group, Inc), intercel‐ lular adhesion molecule (ICAM)1 (1:2,000, cat. no. 60299‐1‐Ig, ProteinTech Group, Inc), syndecan (SDC)4 (1:1,000, cat. no. 11820‐1‐AP, ProteinTech Group, Inc) or β‐actin (1:1,000, cat. no. TA‐09; OriGene Technologies, Inc.).

Techniques: Gene Expression

Journal: Oncology letters

Article Title: Cadherin-16 inhibits thyroid carcinoma cell proliferation and invasion.

doi: 10.3892/ol.2022.13265

Figure Lengend Snippet: Figure 2. CDH16 expression in 35 PTC tissue samples and PTC and follicular cell lines. Expression levels of CDH16 in 35 pairs of PTC and corresponding adjacent N tissue were evaluated by (A) RT‑qPCR and (B) immunohistochemical staining. Scale bar, 25 µm. ***P<0.001 vs. N. CDH16 expression in TC cell lines (BCPAP and TPC1) and the human thyroid follicular cell line Nthy‑ori3‑1 was detected by (C) RT‑qPCR and (D) western blotting. *P<0.05 vs. Nthy‑ori3‑1 cells. Data are presented as the mean ± SD of three independent experiments. N, normal; PTC, papillary thyroid cancer; CDH, cadherin; RT‑q, reverse transcription‑quantitative; TC, thyroid carcinoma.

Article Snippet: The membranes were incubated overnight at 4 ̊C with primary antibodies against CDH16 (1:1,000, cat. no. 15107‐1‐AP, ProteinTech Group, Inc.), DNA polymerase (POL)D1 (1:1,000, cat. no. 15646‐1‐AP, ProteinTech Group, Inc), minichromosome maintenance (MCM)6 (1:2,000, cat. no. 13347‐2‐AP; ProteinTech Group, Inc), claudin (CLDN)1 (1:1,500, cat. no. 13050‐1‐AP, ProteinTech Group, Inc), intercel‐ lular adhesion molecule (ICAM)1 (1:2,000, cat. no. 60299‐1‐Ig, ProteinTech Group, Inc), syndecan (SDC)4 (1:1,000, cat. no. 11820‐1‐AP, ProteinTech Group, Inc) or β‐actin (1:1,000, cat. no. TA‐09; OriGene Technologies, Inc.).

Techniques: Expressing, Immunohistochemical staining, Staining, Western Blot

Journal: Oncology letters

Article Title: Cadherin-16 inhibits thyroid carcinoma cell proliferation and invasion.

doi: 10.3892/ol.2022.13265

Figure Lengend Snippet: Figure 3. GSEA of co‑expressed genes with CDH16 in TC. (A) Volcano map of genes co‑expressed with CDH16 in TCGA samples, as evaluated by LinkedOmics. Red, positively correlated; green, negatively correlated. (B) RNA‑sequencing data from TCGA were analyzed by GSEA enrichment plots for KEGG_DNA_REPLICATION and KEGG_CELL_ADHENSION_MOLECULES_. KEGG, Kyoto Encyclopedia of Genes and Genomes; GSEA, Gene Set Enrichment Analysis; CDH, cadherin.

Article Snippet: The membranes were incubated overnight at 4 ̊C with primary antibodies against CDH16 (1:1,000, cat. no. 15107‐1‐AP, ProteinTech Group, Inc.), DNA polymerase (POL)D1 (1:1,000, cat. no. 15646‐1‐AP, ProteinTech Group, Inc), minichromosome maintenance (MCM)6 (1:2,000, cat. no. 13347‐2‐AP; ProteinTech Group, Inc), claudin (CLDN)1 (1:1,500, cat. no. 13050‐1‐AP, ProteinTech Group, Inc), intercel‐ lular adhesion molecule (ICAM)1 (1:2,000, cat. no. 60299‐1‐Ig, ProteinTech Group, Inc), syndecan (SDC)4 (1:1,000, cat. no. 11820‐1‐AP, ProteinTech Group, Inc) or β‐actin (1:1,000, cat. no. TA‐09; OriGene Technologies, Inc.).

Techniques:

Journal: Oncology letters

Article Title: Cadherin-16 inhibits thyroid carcinoma cell proliferation and invasion.

doi: 10.3892/ol.2022.13265

Figure Lengend Snippet: Figure 4. Correlation between POLD1, MCM6, CLDN1, ICAM1, SDC4 and CDH16 expression was analyzed using the LinkedOmics database. POL, poly‑ merase; MCM, minichromosome maintenance; CLDN, claudin; ICAM, intercellular adhesion molecule; SDC, syndecan; CDH, cadherin.

Article Snippet: The membranes were incubated overnight at 4 ̊C with primary antibodies against CDH16 (1:1,000, cat. no. 15107‐1‐AP, ProteinTech Group, Inc.), DNA polymerase (POL)D1 (1:1,000, cat. no. 15646‐1‐AP, ProteinTech Group, Inc), minichromosome maintenance (MCM)6 (1:2,000, cat. no. 13347‐2‐AP; ProteinTech Group, Inc), claudin (CLDN)1 (1:1,500, cat. no. 13050‐1‐AP, ProteinTech Group, Inc), intercel‐ lular adhesion molecule (ICAM)1 (1:2,000, cat. no. 60299‐1‐Ig, ProteinTech Group, Inc), syndecan (SDC)4 (1:1,000, cat. no. 11820‐1‐AP, ProteinTech Group, Inc) or β‐actin (1:1,000, cat. no. TA‐09; OriGene Technologies, Inc.).

Techniques: Expressing

Journal: Oncology letters

Article Title: Cadherin-16 inhibits thyroid carcinoma cell proliferation and invasion.

doi: 10.3892/ol.2022.13265

Figure Lengend Snippet: Figure 5. CDH16‑Overex inhibits proliferation and promotes apoptosis of BCPAP cells. (A) mRNA levels of CDH16 in CDH16‑Overex compared with control cells were examined by reverse transcription‑quantitative PCR. (B) Protein levels of CDH16 were assessed by western blotting. (C) Cell proliferation was evaluated by MTT assay and (D) apoptosis was evaluated by (E) flow cytometry following transfection. *P<0.05 and ***P<0.001 vs. negative control. Data are presented as the mean ± SD of three independent experiments. n=3; CDH16, cadherin; Overex, overexpression; OD, optical density.

Article Snippet: The membranes were incubated overnight at 4 ̊C with primary antibodies against CDH16 (1:1,000, cat. no. 15107‐1‐AP, ProteinTech Group, Inc.), DNA polymerase (POL)D1 (1:1,000, cat. no. 15646‐1‐AP, ProteinTech Group, Inc), minichromosome maintenance (MCM)6 (1:2,000, cat. no. 13347‐2‐AP; ProteinTech Group, Inc), claudin (CLDN)1 (1:1,500, cat. no. 13050‐1‐AP, ProteinTech Group, Inc), intercel‐ lular adhesion molecule (ICAM)1 (1:2,000, cat. no. 60299‐1‐Ig, ProteinTech Group, Inc), syndecan (SDC)4 (1:1,000, cat. no. 11820‐1‐AP, ProteinTech Group, Inc) or β‐actin (1:1,000, cat. no. TA‐09; OriGene Technologies, Inc.).

Techniques: Control, Western Blot, MTT Assay, Flow Cytometry, Transfection, Negative Control, Over Expression

Journal: Oncology letters

Article Title: Cadherin-16 inhibits thyroid carcinoma cell proliferation and invasion.

doi: 10.3892/ol.2022.13265

Figure Lengend Snippet: Figure 6. Overexpression of CDH16 inhibits migration and invasion of BCPAP cells. (A and B) Cell migration was evaluated by cell scratch test following CDH16 overexpression. Scale bar, 500 µm. (C) Migration ability of BCPAP cells following transfection was determined by (D) Transwell invasion assay. Scale bar, 100 µm. Data are presented as the mean ± SD of three independent experiments. *P<0.05 vs. negative control. CDH, cadherin; Overex, overexpression.

Article Snippet: The membranes were incubated overnight at 4 ̊C with primary antibodies against CDH16 (1:1,000, cat. no. 15107‐1‐AP, ProteinTech Group, Inc.), DNA polymerase (POL)D1 (1:1,000, cat. no. 15646‐1‐AP, ProteinTech Group, Inc), minichromosome maintenance (MCM)6 (1:2,000, cat. no. 13347‐2‐AP; ProteinTech Group, Inc), claudin (CLDN)1 (1:1,500, cat. no. 13050‐1‐AP, ProteinTech Group, Inc), intercel‐ lular adhesion molecule (ICAM)1 (1:2,000, cat. no. 60299‐1‐Ig, ProteinTech Group, Inc), syndecan (SDC)4 (1:1,000, cat. no. 11820‐1‐AP, ProteinTech Group, Inc) or β‐actin (1:1,000, cat. no. TA‐09; OriGene Technologies, Inc.).

Techniques: Over Expression, Migration, Transfection, Transwell Invasion Assay, Negative Control

Journal: Oncology letters

Article Title: Cadherin-16 inhibits thyroid carcinoma cell proliferation and invasion.

doi: 10.3892/ol.2022.13265

Figure Lengend Snippet: Figure 7. Overexpression of CDH16 downregulates DNA replication and cell adhesion pathway genes. (A) Representative western blots showing downregula‑ tion of POLD1, MCM6, CLDN1, ICAM1 and SDC4 following overexpression of CDH16. (B) Relative protein levels were quantified by densitometry of western blots (three replicates). Data are presented as the mean ± SD of three independent experiments. n=3. *P<0.05 vs. negative control. CDH16, cadherin; Overex, overexpression, POL, polymerase; MCM, minichromosome maintenance; CLDN, claudin; ICAM, intercellular adhesion molecule; SDC, syndecan.

Article Snippet: The membranes were incubated overnight at 4 ̊C with primary antibodies against CDH16 (1:1,000, cat. no. 15107‐1‐AP, ProteinTech Group, Inc.), DNA polymerase (POL)D1 (1:1,000, cat. no. 15646‐1‐AP, ProteinTech Group, Inc), minichromosome maintenance (MCM)6 (1:2,000, cat. no. 13347‐2‐AP; ProteinTech Group, Inc), claudin (CLDN)1 (1:1,500, cat. no. 13050‐1‐AP, ProteinTech Group, Inc), intercel‐ lular adhesion molecule (ICAM)1 (1:2,000, cat. no. 60299‐1‐Ig, ProteinTech Group, Inc), syndecan (SDC)4 (1:1,000, cat. no. 11820‐1‐AP, ProteinTech Group, Inc) or β‐actin (1:1,000, cat. no. TA‐09; OriGene Technologies, Inc.).

Techniques: Over Expression, Western Blot, Negative Control