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OriGene human pde6d open reading frame orf myc ddk tagged

In silico analysis of rod-specific photoreceptor cGMP phosphodiesterase <t>(PDE6D)</t> expression in hepatocellular carcinoma (HCC). ( A ) Oncomine TM human cancer microarray database analysis of six patient datasets depicting PDE6D mRNA expression levels in HCCs and non-tumorous livers (* p < 0.05 vs. non-tumorous livers). ( B , C ) Oncomine TM human cancer microarray database analysis of PDE6D expression as detected in large-scale RNA profiling studies comparing diverse carcinomas of different origins (* p < 0.05 vs. average expression).

Human Pde6d Open Reading Frame Orf Myc Ddk Tagged, supplied by OriGene, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/origin+reference+frame+function/PDE6D+(NM_002601)+Human+Tagged+ORF+Clone/pmc06468542-145-13-27
Average 90 stars, based on 2 article reviews

human pde6d open reading frame orf myc ddk tagged - by Bioz Stars, 2026-09

90/100 stars

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1) Product Images from "The Delta Subunit of Rod-Specific Photoreceptor cGMP Phosphodiesterase (PDE6D) Contributes to Hepatocellular Carcinoma Progression"

Article Title: The Delta Subunit of Rod-Specific Photoreceptor cGMP Phosphodiesterase (PDE6D) Contributes to Hepatocellular Carcinoma Progression

Journal: Cancers

doi: 10.3390/cancers11030398

In silico analysis of rod-specific photoreceptor cGMP phosphodiesterase (PDE6D) expression in hepatocellular carcinoma (HCC). ( A ) Oncomine TM human cancer microarray database analysis of six patient datasets depicting PDE6D mRNA expression levels in HCCs and non-tumorous livers (* p < 0.05 vs. non-tumorous livers). ( B , C ) Oncomine TM human cancer microarray database analysis of PDE6D expression as detected in large-scale RNA profiling studies comparing diverse carcinomas of different origins (* p < 0.05 vs. average expression).

Figure Legend Snippet: In silico analysis of rod-specific photoreceptor cGMP phosphodiesterase (PDE6D) expression in hepatocellular carcinoma (HCC). ( A ) Oncomine TM human cancer microarray database analysis of six patient datasets depicting PDE6D mRNA expression levels in HCCs and non-tumorous livers (* p < 0.05 vs. non-tumorous livers). ( B , C ) Oncomine TM human cancer microarray database analysis of PDE6D expression as detected in large-scale RNA profiling studies comparing diverse carcinomas of different origins (* p < 0.05 vs. average expression).

Techniques Used: In Silico, Expressing, Microarray

PDE6D expression in HCC in vivo and in vitro. ( A ) PDE6D mRNA levels as quantified by qRT-PCR analysis of HCC patient samples and paired non-tumorous liver tissues (* p < 0.05). ( B ) Detection of PDE6D mRNA in human HCC cells (PLC, Hep3B, HepG2) after qRT-PCR amplification using gel electrophoresis (left panel) and relative PDE6D mRNA levels (qRT-PCR) in human HCC cell lines (PLC, Hep3B, HepG2) compared with primary human hepatocytes derived from different donors (#1–3) (right panel) (* p < 0.05 vs. hepatocytes). ( C ) Exemplary Western blot image (left panel) and summarized densitometric quantification (right panel) of PDE6D protein levels in HCC cells (PLC, Hep3B, HepG2, Huh-7) compared with hepatocytes derived from different donors (#1–2) (* p < 0.05 vs. hepatocytes).

Figure Legend Snippet: PDE6D expression in HCC in vivo and in vitro. ( A ) PDE6D mRNA levels as quantified by qRT-PCR analysis of HCC patient samples and paired non-tumorous liver tissues (* p < 0.05). ( B ) Detection of PDE6D mRNA in human HCC cells (PLC, Hep3B, HepG2) after qRT-PCR amplification using gel electrophoresis (left panel) and relative PDE6D mRNA levels (qRT-PCR) in human HCC cell lines (PLC, Hep3B, HepG2) compared with primary human hepatocytes derived from different donors (#1–3) (right panel) (* p < 0.05 vs. hepatocytes). ( C ) Exemplary Western blot image (left panel) and summarized densitometric quantification (right panel) of PDE6D protein levels in HCC cells (PLC, Hep3B, HepG2, Huh-7) compared with hepatocytes derived from different donors (#1–2) (* p < 0.05 vs. hepatocytes).

Techniques Used: Expressing, In Vivo, In Vitro, Quantitative RT-PCR, Amplification, Nucleic Acid Electrophoresis, Derivative Assay, Western Blot

Effects of PDE6D knockdown on HCC proliferation and clonogenicity. Prior to functional experiments, HCC cell lines (PLC, Hep3B) were transfected with si-RNA-pools against PDE6D (“PDE6D”) or the according control- si-RNA-pool (“Control”). ( A ) PDE6D mRNA levels as quantified by qRT-PCR analysis (* p < 0.05 vs. control). ( B ) PDE6D protein levels as quantified by Western blot analysis. The left panel depicts an exemplary Western blot image, and the right panel depicts the summarized densitometric quantification (* p < 0.05 vs. control). ( C ) Real-time cell proliferation (xCELLigence). Exemplary proliferation curves (left panel) and quantified “slopes” (summarizing the proliferative ability) (right panel) are shown (* p < 0.05 vs. control). ( D ) Relative (to mean) PDE6D as correlated to CyclinD1 mRNA expression levels (qRT-PCR) in human HCC patient tissue samples. ( E , F ) Anchorage-dependent clonogenic assay (an exemplary image (Hep3B) is depicted in the left panel of ( E )). Quantification of colony number (right panel of ( E )) and sizes ( F ) (* p < 0.05).

Figure Legend Snippet: Effects of PDE6D knockdown on HCC proliferation and clonogenicity. Prior to functional experiments, HCC cell lines (PLC, Hep3B) were transfected with si-RNA-pools against PDE6D (“PDE6D”) or the according control- si-RNA-pool (“Control”). ( A ) PDE6D mRNA levels as quantified by qRT-PCR analysis (* p < 0.05 vs. control). ( B ) PDE6D protein levels as quantified by Western blot analysis. The left panel depicts an exemplary Western blot image, and the right panel depicts the summarized densitometric quantification (* p < 0.05 vs. control). ( C ) Real-time cell proliferation (xCELLigence). Exemplary proliferation curves (left panel) and quantified “slopes” (summarizing the proliferative ability) (right panel) are shown (* p < 0.05 vs. control). ( D ) Relative (to mean) PDE6D as correlated to CyclinD1 mRNA expression levels (qRT-PCR) in human HCC patient tissue samples. ( E , F ) Anchorage-dependent clonogenic assay (an exemplary image (Hep3B) is depicted in the left panel of ( E )). Quantification of colony number (right panel of ( E )) and sizes ( F ) (* p < 0.05).

Techniques Used: Functional Assay, Transfection, Quantitative RT-PCR, Western Blot, Expressing, Clonogenic Assay

Expression and function of PDE6D in sorafenib resistance. ( A ) PDE6D mRNA levels as quantified by qRT-PCR analysis in non-resistant (“non-resist.”) as compared to sorafenib-resistant (“resistant”) Hep3B and HepG2 cell clones (* p < 0.05). ( B ) Exemplary image (left panel) and summarized densitometric quantification (right panel) of Western blot analysis of PDE6D levels in non-resistant (“non-resist.”) as compared to sorafenib-resistant (“resistant”) Hep3B cells (rel DM: relative (PDE6D/Actin) densitometry) (* p < 0.05 vs. non-resist.). ( C , D ) Prior to functional experiments, resistant cells (Hep3B) were transfected with si-RNA-pools against PDE6D (“PDE6D”) or the according control- si-RNA-pool (“Control”). ( C ) Depicts relative proliferation (cell numbers) and ( D ) depicts exemplary clonogenic assays. ( E , F ) Forced overexpression of PDE6D protein (PDE6D-OE) in HCC cells (e.g., PLC) was performed by transfection of a human PDE6D open reading frame (ORF) Myc-DDK-tagged plasmid vector (an empty control vector without the PDE6D ORF was used as control treatment). ( E ) Depicts Western blot analysis depicting the overexpressed Myc-DDK-tagged PDE6D protein after PDE6D-OE as well as endogenous (arrow) PDE6D in both PDE6D-OE and control-treated cells. ( F ) Depicts exemplary images (representing 8 replicate values of 2 independent experiments) of cells cultured in 6-wells for 72 h (100,000 cells were initially seeded per 6-well) and treated with different doses of sorafenib (0, 4, 8 µM).

Figure Legend Snippet: Expression and function of PDE6D in sorafenib resistance. ( A ) PDE6D mRNA levels as quantified by qRT-PCR analysis in non-resistant (“non-resist.”) as compared to sorafenib-resistant (“resistant”) Hep3B and HepG2 cell clones (* p < 0.05). ( B ) Exemplary image (left panel) and summarized densitometric quantification (right panel) of Western blot analysis of PDE6D levels in non-resistant (“non-resist.”) as compared to sorafenib-resistant (“resistant”) Hep3B cells (rel DM: relative (PDE6D/Actin) densitometry) (* p < 0.05 vs. non-resist.). ( C , D ) Prior to functional experiments, resistant cells (Hep3B) were transfected with si-RNA-pools against PDE6D (“PDE6D”) or the according control- si-RNA-pool (“Control”). ( C ) Depicts relative proliferation (cell numbers) and ( D ) depicts exemplary clonogenic assays. ( E , F ) Forced overexpression of PDE6D protein (PDE6D-OE) in HCC cells (e.g., PLC) was performed by transfection of a human PDE6D open reading frame (ORF) Myc-DDK-tagged plasmid vector (an empty control vector without the PDE6D ORF was used as control treatment). ( E ) Depicts Western blot analysis depicting the overexpressed Myc-DDK-tagged PDE6D protein after PDE6D-OE as well as endogenous (arrow) PDE6D in both PDE6D-OE and control-treated cells. ( F ) Depicts exemplary images (representing 8 replicate values of 2 independent experiments) of cells cultured in 6-wells for 72 h (100,000 cells were initially seeded per 6-well) and treated with different doses of sorafenib (0, 4, 8 µM).

Techniques Used: Expressing, Quantitative RT-PCR, Clone Assay, Western Blot, Functional Assay, Transfection, Over Expression, Plasmid Preparation, Cell Culture

TGF-â-mediated regulation of PDE6D and the effect of PDE6D on HCC cell migration. ( A – C ) Quantitative RT-PCR analysis of VIMENTIN, SNAIL, S100A4 ( A ) and KRAS ( B ) mRNA levels in HCC cells (PLC) that were stimulated with different doses of recombinant human TGF-â1 protein for 72–96 hours (* p < 0.05 vs. control). ( C , D ) Quantitative RT-PCR revealing mRNA levels ( C ) as well as protein levels as quantified by Western blot analysis (including a representative Western blot image) (the densitometric values represent two independent Western blot analysis) ( D ) of PDE6D expression in HCC cells (PLC) that were treated with different doses of recombinant human TGF-â1 for 72–96 hours. ( D ) also depicts co-treatment with 15 µM of the TGF-â-receptor-1 (TGFBR1) inhibitor LY2157299 (“galunisertib”) (* p < 0.05 vs. control). ( E , F ) Prior to Boyden chamber experiments, non-resistant HCC cells (PLC, Hep3B) ( E ) and sorafenib-resistant Hep3B cells ( F ) were transfected with si-RNA-pools against PDE6D (“PDE6D”) or the according control- si-RNA-pool (“Control”). Migration (migrating cells per visual field) as measured by Boyden chamber migration assay (duration of migration: 4 hours) is depicted as absolute cell counts ( E ) or as normalized migration ( F ) (* p < 0.05).

Figure Legend Snippet: TGF-â-mediated regulation of PDE6D and the effect of PDE6D on HCC cell migration. ( A – C ) Quantitative RT-PCR analysis of VIMENTIN, SNAIL, S100A4 ( A ) and KRAS ( B ) mRNA levels in HCC cells (PLC) that were stimulated with different doses of recombinant human TGF-â1 protein for 72–96 hours (* p < 0.05 vs. control). ( C , D ) Quantitative RT-PCR revealing mRNA levels ( C ) as well as protein levels as quantified by Western blot analysis (including a representative Western blot image) (the densitometric values represent two independent Western blot analysis) ( D ) of PDE6D expression in HCC cells (PLC) that were treated with different doses of recombinant human TGF-â1 for 72–96 hours. ( D ) also depicts co-treatment with 15 µM of the TGF-â-receptor-1 (TGFBR1) inhibitor LY2157299 (“galunisertib”) (* p < 0.05 vs. control). ( E , F ) Prior to Boyden chamber experiments, non-resistant HCC cells (PLC, Hep3B) ( E ) and sorafenib-resistant Hep3B cells ( F ) were transfected with si-RNA-pools against PDE6D (“PDE6D”) or the according control- si-RNA-pool (“Control”). Migration (migrating cells per visual field) as measured by Boyden chamber migration assay (duration of migration: 4 hours) is depicted as absolute cell counts ( E ) or as normalized migration ( F ) (* p < 0.05).

Techniques Used: Migration, Quantitative RT-PCR, Recombinant, Western Blot, Expressing, Transfection

$Expression and cellular localization of PDE6D in HCC in vivo and in vitro. ( A ) PDE6D staining (exemplary images) of non-tumorous liver tissues (left side) and HCC tissues (right side) deposited on the Human Protein Atlas database. ( B ) Tissue microarray analysis of PDE6D expression levels in human HCC tissues ( N = 117) as compared with corresponding non-tumorous liver tissues ( N = 127) (Fisher’s exact P < 0.001). ( C , D ) Tissue microarray analysis of PDE6D expression levels in human HCC tissues correlated with tumor grading (Fisher’s exact P = 0.030) ( C ) and tumor stages ( D ). ( E ) Exemplary immunohistological images of PDE6D protein expression in human HCC samples and corresponding non-tumorous liver tissues applying a tissue microarray revealing nuclear staining next to cytoplasmatic staining patterns (paired samples of two different patients (#1, #2) are depicted). ( F , G ) Tissue microarray analysis comparing tumor stages ( F ) and ERK activation (p-ERK) ( G ) in human HCC tissues with (“yes”) and without (“no”) cytoplasmatic localization pattern of PDE6D. ( H ) In silico-based analysis of importin-á-dependent nuclear localization signals (NLS, red letters ) using the “cNLS Mapper” predicted bipartite NLS in both isoforms of PDE6D (score for both isoforms was 5.3). A legend depicts that higher scores indicate stronger NLS activity and defines major localizations in dependence of each score. ( I , J ) Exemplary immunofluorescence ( I ) and Western blot analysis ( J ) depicting nuclear localization of PDE6D (I) and expression of PDE6D in both nuclear and cytoplasmatic fractions ( J ) of HCC cell lysates.$
Figure Legend Snippet: Expression and cellular localization of PDE6D in HCC in vivo and in vitro. ( A ) PDE6D staining (exemplary images) of non-tumorous liver tissues (left side) and HCC tissues (right side) deposited on the Human Protein Atlas database. ( B ) Tissue microarray analysis of PDE6D expression levels in human HCC tissues ( N = 117) as compared with corresponding non-tumorous liver tissues ( N = 127) (Fisher’s exact P < 0.001). ( C , D ) Tissue microarray analysis of PDE6D expression levels in human HCC tissues correlated with tumor grading (Fisher’s exact P = 0.030) ( C ) and tumor stages ( D ). ( E ) Exemplary immunohistological images of PDE6D protein expression in human HCC samples and corresponding non-tumorous liver tissues applying a tissue microarray revealing nuclear staining next to cytoplasmatic staining patterns (paired samples of two different patients (#1, #2) are depicted). ( F , G ) Tissue microarray analysis comparing tumor stages ( F ) and ERK activation (p-ERK) ( G ) in human HCC tissues with (“yes”) and without (“no”) cytoplasmatic localization pattern of PDE6D. ( H ) In silico-based analysis of importin-á-dependent nuclear localization signals (NLS, red letters ) using the “cNLS Mapper” predicted bipartite NLS in both isoforms of PDE6D (score for both isoforms was 5.3). A legend depicts that higher scores indicate stronger NLS activity and defines major localizations in dependence of each score. ( I , J ) Exemplary immunofluorescence ( I ) and Western blot analysis ( J ) depicting nuclear localization of PDE6D (I) and expression of PDE6D in both nuclear and cytoplasmatic fractions ( J ) of HCC cell lysates.

Techniques Used: Expressing, In Vivo, In Vitro, Staining, Microarray, Activation Assay, In Silico, Activity Assay, Immunofluorescence, Western Blot

Over Expression:

Article Title: The Delta Subunit of Rod-Specific Photoreceptor cGMP Phosphodiesterase (PDE6D) Contributes to Hepatocellular Carcinoma Progression
Article Snippet: For siRNA-induced knockdown of PDE6D, a “ si-RNA-pool -PDE6D” (functionally verified by siTOOLs Biotech GmbH, Planegg, Germany) was used [ ]. .. Overexpression of PDE6D protein in HCC cells was induced by transfection of a human PDE6D open reading frame (ORF) Myc-DDK-tagged plasmid vector (pCMV6-Entry cDNA vector system) from OriGene (Rockville, MD, USA) (CAT#: RC203172) (an according empty control vector without the PDE6D ORF was used as control treatment). ..

Transfection:

Plasmid Preparation:

Control:

Construct:

Article Title: A homozygous PDE6D mutation in Joubert syndrome impairs targeting of farnesylated INPP5E protein to the primary cilium
Article Snippet: .. DNA constructs C-terminal Myc-DDK-tagged ORF clone of PDE6D ( {"type":"entrez-nucleotide","attrs":{"text":"NM_002601.2","term_id":"56676309","term_text":"NM_002601.2"}} NM_002601.2 ) was obtained from Origene (RC203172). .. Deletion of exon 3 was performed using the QuikChange site-directed mutagenesis kit (Stratagene) according to the manufacturer’s instructions and were confirmed by direct sequencing (Primers used are listed in Supp.

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