primary human dpcs Search Results


human primary dental pulp cells dpc  (ATCC)
99
ATCC human primary dental pulp cells dpc
M. hyorhinis promotes TNF-α secretion from PCa cells. (A), relative TNF-α secretion levels in M. hyorhinis-contaminated PCa cells (PC3-cM and C4–2B-cM) and mycoplasma-free cells; multiple cancer cells (PC3, C4–2B, DU145, LNCaP, MDA-MB-231, MCF-7, MCF10A, HeLa, MG-63), HDMEC, HF, and <t>DPC</t> via ELISA. (B), experimental design. The parental PCa cells (PC3–P and C4–2B–P) were infected with M. hyorhinis (3 × 107 CFU, 5 MOI) for 2 passages and then passaged twice a week for 6 weeks without further infection (PC3-M and C4–2B-M). For PC3-MF and C4–2B-MF cells, 4 weeks after infection, M. hyorhinis was eliminated in cell cultures for 3 weeks. Seven weeks after infection, in vitro assays and analyses were performed using these PCa cells. (C), relative TNF-α secretion levels in the parental (PC3–P and C4–2B–P) and M. hyorhinis-infected (PC3-M and C4–2B-M) PCa cells. (D), relative TNF-α secretion levels in PC3-M and C4–2B-M and previously infected PCa cells after elimination of M. hyorhinis (PC3-MF and C4–2B-MF). M. hyrorhinis infection was confirmed by Western blotting using specific anti-M. hyorhinis (P70 surface antigen) antibody. βactin was used as a loading control. See full images in Supplementary Figure S4. All results represent mean ± SD values from triplicate assays, and the experiments were repeated three times. **p < 0.0001. HDMEC, human dermal microvascular endothelial cells; HF, human <t>primary</t> <t>fibroblasts;</t> DPC, human primary dental pulp cells; CFU, colony-forming units; MOI, multiplicity of infection.
Human Primary Dental Pulp Cells Dpc, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human hair follicle dpcs hfdpcs  (PromoCell)
95
PromoCell human hair follicle dpcs hfdpcs
M. hyorhinis promotes TNF-α secretion from PCa cells. (A), relative TNF-α secretion levels in M. hyorhinis-contaminated PCa cells (PC3-cM and C4–2B-cM) and mycoplasma-free cells; multiple cancer cells (PC3, C4–2B, DU145, LNCaP, MDA-MB-231, MCF-7, MCF10A, HeLa, MG-63), HDMEC, HF, and <t>DPC</t> via ELISA. (B), experimental design. The parental PCa cells (PC3–P and C4–2B–P) were infected with M. hyorhinis (3 × 107 CFU, 5 MOI) for 2 passages and then passaged twice a week for 6 weeks without further infection (PC3-M and C4–2B-M). For PC3-MF and C4–2B-MF cells, 4 weeks after infection, M. hyorhinis was eliminated in cell cultures for 3 weeks. Seven weeks after infection, in vitro assays and analyses were performed using these PCa cells. (C), relative TNF-α secretion levels in the parental (PC3–P and C4–2B–P) and M. hyorhinis-infected (PC3-M and C4–2B-M) PCa cells. (D), relative TNF-α secretion levels in PC3-M and C4–2B-M and previously infected PCa cells after elimination of M. hyorhinis (PC3-MF and C4–2B-MF). M. hyrorhinis infection was confirmed by Western blotting using specific anti-M. hyorhinis (P70 surface antigen) antibody. βactin was used as a loading control. See full images in Supplementary Figure S4. All results represent mean ± SD values from triplicate assays, and the experiments were repeated three times. **p < 0.0001. HDMEC, human dermal microvascular endothelial cells; HF, human <t>primary</t> <t>fibroblasts;</t> DPC, human primary dental pulp cells; CFU, colony-forming units; MOI, multiplicity of infection.
Human Hair Follicle Dpcs Hfdpcs, supplied by PromoCell, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dpc4  (Santa Cruz Biotechnology)
96
Santa Cruz Biotechnology dpc4
M. hyorhinis promotes TNF-α secretion from PCa cells. (A), relative TNF-α secretion levels in M. hyorhinis-contaminated PCa cells (PC3-cM and C4–2B-cM) and mycoplasma-free cells; multiple cancer cells (PC3, C4–2B, DU145, LNCaP, MDA-MB-231, MCF-7, MCF10A, HeLa, MG-63), HDMEC, HF, and <t>DPC</t> via ELISA. (B), experimental design. The parental PCa cells (PC3–P and C4–2B–P) were infected with M. hyorhinis (3 × 107 CFU, 5 MOI) for 2 passages and then passaged twice a week for 6 weeks without further infection (PC3-M and C4–2B-M). For PC3-MF and C4–2B-MF cells, 4 weeks after infection, M. hyorhinis was eliminated in cell cultures for 3 weeks. Seven weeks after infection, in vitro assays and analyses were performed using these PCa cells. (C), relative TNF-α secretion levels in the parental (PC3–P and C4–2B–P) and M. hyorhinis-infected (PC3-M and C4–2B-M) PCa cells. (D), relative TNF-α secretion levels in PC3-M and C4–2B-M and previously infected PCa cells after elimination of M. hyorhinis (PC3-MF and C4–2B-MF). M. hyrorhinis infection was confirmed by Western blotting using specific anti-M. hyorhinis (P70 surface antigen) antibody. βactin was used as a loading control. See full images in Supplementary Figure S4. All results represent mean ± SD values from triplicate assays, and the experiments were repeated three times. **p < 0.0001. HDMEC, human dermal microvascular endothelial cells; HF, human <t>primary</t> <t>fibroblasts;</t> DPC, human primary dental pulp cells; CFU, colony-forming units; MOI, multiplicity of infection.
Dpc4, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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primary human hair dpcs (hdpcs)  (ScienCell)
90
ScienCell primary human hair dpcs (hdpcs)
The Cinnamomum osmophloeum Kanehira extract enhances the proliferation of cultured rat vibrissae dermal papilla cells <t>(DPCs).</t> Cell proliferation was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Mino: minoxidil. Values are the mean ± SD . * P < 0.05 and ** P < 0.01, compared with untreated cells (control).
Primary Human Hair Dpcs (Hdpcs), supplied by ScienCell, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human smad4/dpc4 cdna  (Johns Hopkins HealthCare)
90
Johns Hopkins HealthCare human smad4/dpc4 cdna
The Cinnamomum osmophloeum Kanehira extract enhances the proliferation of cultured rat vibrissae dermal papilla cells <t>(DPCs).</t> Cell proliferation was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Mino: minoxidil. Values are the mean ± SD . * P < 0.05 and ** P < 0.01, compared with untreated cells (control).
Human Smad4/Dpc4 Cdna, supplied by Johns Hopkins HealthCare, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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gene exp dpcd hs00381790 m1  (Thermo Fisher)
92
Thermo Fisher gene exp dpcd hs00381790 m1
Figure 1. Discovery of <t>DPCD</t> through R2TP BioID (A) Overlap of interactions identified in this study and previously reported interactions from BioGrid (Biological General Repository for Interaction Datasets).33
Gene Exp Dpcd Hs00381790 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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primary human dpcs  (PromoCell)
95
PromoCell primary human dpcs
Figure 1. Discovery of <t>DPCD</t> through R2TP BioID (A) Overlap of interactions identified in this study and previously reported interactions from BioGrid (Biological General Repository for Interaction Datasets).33
Primary Human Dpcs, supplied by PromoCell, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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antibodies for smad4  (Proteintech)
96
Proteintech antibodies for smad4
Figure 1. Discovery of <t>DPCD</t> through R2TP BioID (A) Overlap of interactions identified in this study and previously reported interactions from BioGrid (Biological General Repository for Interaction Datasets).33
Antibodies For Smad4, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti-human type ii collagen antibody  (DPC Biermann GmbH)
90
DPC Biermann GmbH rabbit anti-human type ii collagen antibody
Figure 1. Discovery of <t>DPCD</t> through R2TP BioID (A) Overlap of interactions identified in this study and previously reported interactions from BioGrid (Biological General Repository for Interaction Datasets).33
Rabbit Anti Human Type Ii Collagen Antibody, supplied by DPC Biermann GmbH, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dpcr1 polyclonal antibody, abby fluor 555 conjugated  (Bioss)
N/A
Making up nearly 6% of the human genome, chromosome 6 contains around 1,200 genes within 170 million base pairs of sequence. Deletion of a portion of the q arm of chromosome 6 is associated with
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dpcr1 polyclonal antibody, apc-cy5.5 conjugated  (Bioss)
N/A
Making up nearly 6% of the human genome, chromosome 6 contains around 1,200 genes within 170 million base pairs of sequence. Deletion of a portion of the q arm of chromosome 6 is associated with
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dpcr1 polyclonal antibody, pe-cy3 conjugated  (Bioss)
N/A
Making up nearly 6% of the human genome, chromosome 6 contains around 1,200 genes within 170 million base pairs of sequence. Deletion of a portion of the q arm of chromosome 6 is associated with
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Image Search Results


M. hyorhinis promotes TNF-α secretion from PCa cells. (A), relative TNF-α secretion levels in M. hyorhinis-contaminated PCa cells (PC3-cM and C4–2B-cM) and mycoplasma-free cells; multiple cancer cells (PC3, C4–2B, DU145, LNCaP, MDA-MB-231, MCF-7, MCF10A, HeLa, MG-63), HDMEC, HF, and DPC via ELISA. (B), experimental design. The parental PCa cells (PC3–P and C4–2B–P) were infected with M. hyorhinis (3 × 107 CFU, 5 MOI) for 2 passages and then passaged twice a week for 6 weeks without further infection (PC3-M and C4–2B-M). For PC3-MF and C4–2B-MF cells, 4 weeks after infection, M. hyorhinis was eliminated in cell cultures for 3 weeks. Seven weeks after infection, in vitro assays and analyses were performed using these PCa cells. (C), relative TNF-α secretion levels in the parental (PC3–P and C4–2B–P) and M. hyorhinis-infected (PC3-M and C4–2B-M) PCa cells. (D), relative TNF-α secretion levels in PC3-M and C4–2B-M and previously infected PCa cells after elimination of M. hyorhinis (PC3-MF and C4–2B-MF). M. hyrorhinis infection was confirmed by Western blotting using specific anti-M. hyorhinis (P70 surface antigen) antibody. βactin was used as a loading control. See full images in Supplementary Figure S4. All results represent mean ± SD values from triplicate assays, and the experiments were repeated three times. **p < 0.0001. HDMEC, human dermal microvascular endothelial cells; HF, human primary fibroblasts; DPC, human primary dental pulp cells; CFU, colony-forming units; MOI, multiplicity of infection.

Journal: Heliyon

Article Title: Mycoplasma hyorhinis infection promotes TNF-α signaling and SMAC mimetic-mediated apoptosis in human prostate cancer

doi: 10.1016/j.heliyon.2023.e20655

Figure Lengend Snippet: M. hyorhinis promotes TNF-α secretion from PCa cells. (A), relative TNF-α secretion levels in M. hyorhinis-contaminated PCa cells (PC3-cM and C4–2B-cM) and mycoplasma-free cells; multiple cancer cells (PC3, C4–2B, DU145, LNCaP, MDA-MB-231, MCF-7, MCF10A, HeLa, MG-63), HDMEC, HF, and DPC via ELISA. (B), experimental design. The parental PCa cells (PC3–P and C4–2B–P) were infected with M. hyorhinis (3 × 107 CFU, 5 MOI) for 2 passages and then passaged twice a week for 6 weeks without further infection (PC3-M and C4–2B-M). For PC3-MF and C4–2B-MF cells, 4 weeks after infection, M. hyorhinis was eliminated in cell cultures for 3 weeks. Seven weeks after infection, in vitro assays and analyses were performed using these PCa cells. (C), relative TNF-α secretion levels in the parental (PC3–P and C4–2B–P) and M. hyorhinis-infected (PC3-M and C4–2B-M) PCa cells. (D), relative TNF-α secretion levels in PC3-M and C4–2B-M and previously infected PCa cells after elimination of M. hyorhinis (PC3-MF and C4–2B-MF). M. hyrorhinis infection was confirmed by Western blotting using specific anti-M. hyorhinis (P70 surface antigen) antibody. βactin was used as a loading control. See full images in Supplementary Figure S4. All results represent mean ± SD values from triplicate assays, and the experiments were repeated three times. **p < 0.0001. HDMEC, human dermal microvascular endothelial cells; HF, human primary fibroblasts; DPC, human primary dental pulp cells; CFU, colony-forming units; MOI, multiplicity of infection.

Article Snippet: Human primary fibroblasts (HF) [ ], human primary dental pulp cells (DPC) [ ], and human cancer cell lines (PC3, C4–2B, DU145, LNCaP, MDA-MB-231, MCF-7, MCF10A, HeLa, MG-63; American Type Culture Collection (ATCC), Manassas, VA) were grown in Dulbecco's modified Eagle's medium (DMEM, Invitrogen, Carsbad, CA) supplemented with 10 % FBS and 1 % penicillin/streptomycin.

Techniques: Enzyme-linked Immunosorbent Assay, Infection, In Vitro, Western Blot

The Cinnamomum osmophloeum Kanehira extract enhances the proliferation of cultured rat vibrissae dermal papilla cells (DPCs). Cell proliferation was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Mino: minoxidil. Values are the mean ± SD . * P < 0.05 and ** P < 0.01, compared with untreated cells (control).

Journal: Cell Transplantation

Article Title: Effect of Cinnamomum osmophloeum Kanehira Leaf Aqueous Extract on Dermal Papilla Cell Proliferation and Hair Growth

doi: 10.1177/0963689717741139

Figure Lengend Snippet: The Cinnamomum osmophloeum Kanehira extract enhances the proliferation of cultured rat vibrissae dermal papilla cells (DPCs). Cell proliferation was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Mino: minoxidil. Values are the mean ± SD . * P < 0.05 and ** P < 0.01, compared with untreated cells (control).

Article Snippet: Primary human hair DPCs (hDPCs) were purchased from ScienCell Research Laboratories (Carlsbad, CA, USA) as primary cells and maintained in mesenchymal stem cell (MSC) medium (ScienCell Research Laboratories), 5% (w/v) FBS, 1% (w/v) penicillin, 1% (w/v) mesangial cell growth supplement (ScienCell Research Laboratories) under a 5% CO , humidified environment at 37 °C.

Techniques: Cell Culture, MTT Assay, Control

The Cinnamomum osmophloeum Kanehira extract enhances the proliferation of cultured human hair dermal papilla cells (hDPCs). Cell proliferation was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Mino: minoxidil. Values are the mean ± SD . * P < 0.05 and ** P < 0.01, compared with untreated cells (control).

Journal: Cell Transplantation

Article Title: Effect of Cinnamomum osmophloeum Kanehira Leaf Aqueous Extract on Dermal Papilla Cell Proliferation and Hair Growth

doi: 10.1177/0963689717741139

Figure Lengend Snippet: The Cinnamomum osmophloeum Kanehira extract enhances the proliferation of cultured human hair dermal papilla cells (hDPCs). Cell proliferation was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Mino: minoxidil. Values are the mean ± SD . * P < 0.05 and ** P < 0.01, compared with untreated cells (control).

Article Snippet: Primary human hair DPCs (hDPCs) were purchased from ScienCell Research Laboratories (Carlsbad, CA, USA) as primary cells and maintained in mesenchymal stem cell (MSC) medium (ScienCell Research Laboratories), 5% (w/v) FBS, 1% (w/v) penicillin, 1% (w/v) mesangial cell growth supplement (ScienCell Research Laboratories) under a 5% CO , humidified environment at 37 °C.

Techniques: Cell Culture, MTT Assay, Control

Quantitative polymerase chain reaction of growth factor genes in human hair dermal papilla cells (hDPCs). Changes in growth factor messenger RNA (mRNA) expression levels induced after different treatments for 48 h. The growth factor mRNA expression levels were normalized to that of β-actin mRNA expression with the results expressed as fold changes.

Journal: Cell Transplantation

Article Title: Effect of Cinnamomum osmophloeum Kanehira Leaf Aqueous Extract on Dermal Papilla Cell Proliferation and Hair Growth

doi: 10.1177/0963689717741139

Figure Lengend Snippet: Quantitative polymerase chain reaction of growth factor genes in human hair dermal papilla cells (hDPCs). Changes in growth factor messenger RNA (mRNA) expression levels induced after different treatments for 48 h. The growth factor mRNA expression levels were normalized to that of β-actin mRNA expression with the results expressed as fold changes.

Article Snippet: Primary human hair DPCs (hDPCs) were purchased from ScienCell Research Laboratories (Carlsbad, CA, USA) as primary cells and maintained in mesenchymal stem cell (MSC) medium (ScienCell Research Laboratories), 5% (w/v) FBS, 1% (w/v) penicillin, 1% (w/v) mesangial cell growth supplement (ScienCell Research Laboratories) under a 5% CO , humidified environment at 37 °C.

Techniques: Real-time Polymerase Chain Reaction, Expressing

Effect of Cinnamomum osmophloeum Kanehira (COK) extract and minoxidil on the hair follicle unit area and skin thickness of mice. (a) Biopsied dorsal skin of C57BL/6 mice at days 1 and 14 was fixed and then stained with hematoxylin and eosin. Effects of topical COK extracts on C57BL/6 mouse dermal papilla cells (DPCs) during the anagen phase induction shown by representative skin samples from 4 animals. (b) Quantified slides from these samples. Scale bars = 200 µm.

Journal: Cell Transplantation

Article Title: Effect of Cinnamomum osmophloeum Kanehira Leaf Aqueous Extract on Dermal Papilla Cell Proliferation and Hair Growth

doi: 10.1177/0963689717741139

Figure Lengend Snippet: Effect of Cinnamomum osmophloeum Kanehira (COK) extract and minoxidil on the hair follicle unit area and skin thickness of mice. (a) Biopsied dorsal skin of C57BL/6 mice at days 1 and 14 was fixed and then stained with hematoxylin and eosin. Effects of topical COK extracts on C57BL/6 mouse dermal papilla cells (DPCs) during the anagen phase induction shown by representative skin samples from 4 animals. (b) Quantified slides from these samples. Scale bars = 200 µm.

Article Snippet: Primary human hair DPCs (hDPCs) were purchased from ScienCell Research Laboratories (Carlsbad, CA, USA) as primary cells and maintained in mesenchymal stem cell (MSC) medium (ScienCell Research Laboratories), 5% (w/v) FBS, 1% (w/v) penicillin, 1% (w/v) mesangial cell growth supplement (ScienCell Research Laboratories) under a 5% CO , humidified environment at 37 °C.

Techniques: Staining

Figure 1. Discovery of DPCD through R2TP BioID (A) Overlap of interactions identified in this study and previously reported interactions from BioGrid (Biological General Repository for Interaction Datasets).33

Journal: Cell reports

Article Title: DPCD is a regulator of R2TP in ciliogenesis initiation through Akt signaling.

doi: 10.1016/j.celrep.2024.113713

Figure Lengend Snippet: Figure 1. Discovery of DPCD through R2TP BioID (A) Overlap of interactions identified in this study and previously reported interactions from BioGrid (Biological General Repository for Interaction Datasets).33

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Hygromycin B Thermo Fisher Scientific 10687010; CAS: 31282-04-9 Tetracycline Bioshop TET701; CAS: 64-75-5 Biotin Bioshop BIO302; CAS: 58-85-5 Paraformaldehyde 32% Aqueous Solution EM Grade Electron Microscopy Sciences 15714; CAS: 30525-89-4 Bovine serum albumin Bioshop ALB001; CAS: 30525-89-4 Puromycin Bioshop PUR333; CAS: 58-58-2 JG-98 Cayman Chemicals 38337; CAS: 1456551-16-8 Geldanamycin Cayman Chemicals 13355; CAS: 30562-34-6 MK-2206 Cayman Chemicals 15569; CAS: 681281-88-9 CB-6644 Cayman Chemicals 36125; CAS: 2316817-88-4 AKTiIV (Akt Inhibitor IV) Cayman Chemicals 11593; CAS: 1032350-13-2 SIGMAFAST Protease inhibitor Tablets Sigma S8820 3x-FLAG Peptide Sigma F4799 AKT2, Active, His-tag Recombinant (phosphorylated) BPS Bioscience 40011 AKT2, Inactive, His-tag Recombinant BPS Bioscience 40001 Critical commercial assays PureLink RNA Mini Kit Thermo Fisher Scientific 12183018A QIAGEN OneStep RT-PCR Kit Qiagen 210215 Deposited data BioID mass spectrometry data This paper, deposited in MassIVE MSV000091230 DPCD SAXS model SASBDB SASDQ98 Experimental models: Cell lines HEK293 Flp-In T-REX Dr. Liliana Attisano, University of Toronto N/A HEK293T Dr. Liliana Attisano, University of Toronto N/A hTERT RPE-1 Dr. Laurence Pelletier, Lunenfeld- Tanenbaum Research Institute N/A RPE-1 DPCD KO This paper N/A RPE-1 RPAP3 KO This paper N/A RPE-1 PIH1D1 KO This paper N/A Oligonucleotides siGENOME Control Pool Non-Targeting #1 Dharmacon D-001206-13-05 siGENOME SMARTpool Human RUVBL1 Dharmacon M-008977-01-0005 siGENOME SMARTpool Human RUVBL2 Dharmacon M-012299-00-0005 siGENOME SMARTpool Human PIH1D1 Dharmacon M-020963-01-005 siGENOME SMARTpool Human DPCD Dharmacon M-027328-00-005 ON-TARGETplus SMARTpool Human RPAP3 Dharmacon L-014385-01-005 DPCD Taqman probe Thermo Fisher Scientific 4331182; assay ID: Hs00381790_m1 GAPDH Taqman probe Thermo Fisher Scientific 4448489; assay ID: Hs02786624_g1 5’-TTCAAGCTGCCACTGGC-3’ This paper DPCD sgRNA 5’-CAGCAGCTTCGGGTTCG-3’ This paper PIH1D1 sgRNA 5’-AGCACAACAGAATAAGCAGC-3’ Hart et al.77 RPAP3 sgRNA Recombinant DNA See Table S4 for list of recombinant DNA See Table S4 N/A (Continued on next page) Cell Reports 43, 113713, February 27, 2024 19

Techniques:

Figure 3. R2TP and DPCD regulate ciliogenesis through Akt signaling and Rabin8 pre-ciliary trafficking (A) Model for R2TP-DPCD regulation of Akt signaling and ciliogenesis. (B) Top: representative western blots of RPE-1 cells after siRNA knockdown for 72 h and serum starvation for 3 h. Bottom: corresponding quantification of relative p-Akt levels normalized against Akt and GAPDH. (C) Representative western blots of WT and KO RPE-1 cells serum starved for 3 h, immunoblotted (top), and quantified (bottom) as described in (B). (D) Top: western blot analysis of lysates from RPE-1 KO cells transfected with empty vectors or vectors expressing indicated proteins. Bottom: corresponding quantifications as described in (B).

Journal: Cell reports

Article Title: DPCD is a regulator of R2TP in ciliogenesis initiation through Akt signaling.

doi: 10.1016/j.celrep.2024.113713

Figure Lengend Snippet: Figure 3. R2TP and DPCD regulate ciliogenesis through Akt signaling and Rabin8 pre-ciliary trafficking (A) Model for R2TP-DPCD regulation of Akt signaling and ciliogenesis. (B) Top: representative western blots of RPE-1 cells after siRNA knockdown for 72 h and serum starvation for 3 h. Bottom: corresponding quantification of relative p-Akt levels normalized against Akt and GAPDH. (C) Representative western blots of WT and KO RPE-1 cells serum starved for 3 h, immunoblotted (top), and quantified (bottom) as described in (B). (D) Top: western blot analysis of lysates from RPE-1 KO cells transfected with empty vectors or vectors expressing indicated proteins. Bottom: corresponding quantifications as described in (B).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Hygromycin B Thermo Fisher Scientific 10687010; CAS: 31282-04-9 Tetracycline Bioshop TET701; CAS: 64-75-5 Biotin Bioshop BIO302; CAS: 58-85-5 Paraformaldehyde 32% Aqueous Solution EM Grade Electron Microscopy Sciences 15714; CAS: 30525-89-4 Bovine serum albumin Bioshop ALB001; CAS: 30525-89-4 Puromycin Bioshop PUR333; CAS: 58-58-2 JG-98 Cayman Chemicals 38337; CAS: 1456551-16-8 Geldanamycin Cayman Chemicals 13355; CAS: 30562-34-6 MK-2206 Cayman Chemicals 15569; CAS: 681281-88-9 CB-6644 Cayman Chemicals 36125; CAS: 2316817-88-4 AKTiIV (Akt Inhibitor IV) Cayman Chemicals 11593; CAS: 1032350-13-2 SIGMAFAST Protease inhibitor Tablets Sigma S8820 3x-FLAG Peptide Sigma F4799 AKT2, Active, His-tag Recombinant (phosphorylated) BPS Bioscience 40011 AKT2, Inactive, His-tag Recombinant BPS Bioscience 40001 Critical commercial assays PureLink RNA Mini Kit Thermo Fisher Scientific 12183018A QIAGEN OneStep RT-PCR Kit Qiagen 210215 Deposited data BioID mass spectrometry data This paper, deposited in MassIVE MSV000091230 DPCD SAXS model SASBDB SASDQ98 Experimental models: Cell lines HEK293 Flp-In T-REX Dr. Liliana Attisano, University of Toronto N/A HEK293T Dr. Liliana Attisano, University of Toronto N/A hTERT RPE-1 Dr. Laurence Pelletier, Lunenfeld- Tanenbaum Research Institute N/A RPE-1 DPCD KO This paper N/A RPE-1 RPAP3 KO This paper N/A RPE-1 PIH1D1 KO This paper N/A Oligonucleotides siGENOME Control Pool Non-Targeting #1 Dharmacon D-001206-13-05 siGENOME SMARTpool Human RUVBL1 Dharmacon M-008977-01-0005 siGENOME SMARTpool Human RUVBL2 Dharmacon M-012299-00-0005 siGENOME SMARTpool Human PIH1D1 Dharmacon M-020963-01-005 siGENOME SMARTpool Human DPCD Dharmacon M-027328-00-005 ON-TARGETplus SMARTpool Human RPAP3 Dharmacon L-014385-01-005 DPCD Taqman probe Thermo Fisher Scientific 4331182; assay ID: Hs00381790_m1 GAPDH Taqman probe Thermo Fisher Scientific 4448489; assay ID: Hs02786624_g1 5’-TTCAAGCTGCCACTGGC-3’ This paper DPCD sgRNA 5’-CAGCAGCTTCGGGTTCG-3’ This paper PIH1D1 sgRNA 5’-AGCACAACAGAATAAGCAGC-3’ Hart et al.77 RPAP3 sgRNA Recombinant DNA See Table S4 for list of recombinant DNA See Table S4 N/A (Continued on next page) Cell Reports 43, 113713, February 27, 2024 19

Techniques: Western Blot, Knockdown, Transfection, Expressing

Figure 5. Mapping the interaction interfaces between DPCD and RUVBL1/2 (A) His-SUMO-DPCD was used to pull down RUVBL complexes in which one of the subunits lacked the domain II external (DIIext). R1 and R2 refer to RUVBL1 and RUVBL2, respectively. A degradation band of RUVBL1 is indicated by *. (B) His-SUMO-DPCD truncated constructs were used to pull down RUVBL1/2 (left) and RUVBL2 (right). The length of each construct is indicated at the top of each lane. His-SUMO was used as negative control. FL, full length. (C) Integration of pull-down results shown in (B) with sequence conservation analysis of DPCD. On the left, the region where RUVBL1/2 was found to bind on DPCD is indicated (dashed square). DPCD domains 1 and 2 (DD1 and DD2) and the linker are colored in gray, red, and green, respectively. On the right, the conservation of the region between amino acids 100 and 180 of DPCD is shown as logo. A highly conserved sequence (amino acids 133–140) was identified and is highlighted in teal blue. Chemical properties of amino acids in the logo are indicated by colors. (D) Schematics of DPCD mutants made based on the sequence conservation analysis in (C). For simplicity, the name of each mutant used throughout this work is according to this scheme. (E) His-SUMO-DPCD mutants were used to pull down RUVBL1/2 (top) and RUVBL2 (bottom). All pull-down experiments were performed in parallel and with the same concentration of proteins for comparison. His-SUMO was used as bait in negative controls. (F) Top: effect of DPCD mutants on ciliogenesis in RPE-1 DPCD-KO cells. RPE-1 cells expressing endogenous DPCD and transfected with empty vector (EV, black) were used as control. RPE-1 DPCD-KO cells were transfected with EV (black), WT DPCD, or DPCD mutant-expressing plasmids (highlighted in red).

Journal: Cell reports

Article Title: DPCD is a regulator of R2TP in ciliogenesis initiation through Akt signaling.

doi: 10.1016/j.celrep.2024.113713

Figure Lengend Snippet: Figure 5. Mapping the interaction interfaces between DPCD and RUVBL1/2 (A) His-SUMO-DPCD was used to pull down RUVBL complexes in which one of the subunits lacked the domain II external (DIIext). R1 and R2 refer to RUVBL1 and RUVBL2, respectively. A degradation band of RUVBL1 is indicated by *. (B) His-SUMO-DPCD truncated constructs were used to pull down RUVBL1/2 (left) and RUVBL2 (right). The length of each construct is indicated at the top of each lane. His-SUMO was used as negative control. FL, full length. (C) Integration of pull-down results shown in (B) with sequence conservation analysis of DPCD. On the left, the region where RUVBL1/2 was found to bind on DPCD is indicated (dashed square). DPCD domains 1 and 2 (DD1 and DD2) and the linker are colored in gray, red, and green, respectively. On the right, the conservation of the region between amino acids 100 and 180 of DPCD is shown as logo. A highly conserved sequence (amino acids 133–140) was identified and is highlighted in teal blue. Chemical properties of amino acids in the logo are indicated by colors. (D) Schematics of DPCD mutants made based on the sequence conservation analysis in (C). For simplicity, the name of each mutant used throughout this work is according to this scheme. (E) His-SUMO-DPCD mutants were used to pull down RUVBL1/2 (top) and RUVBL2 (bottom). All pull-down experiments were performed in parallel and with the same concentration of proteins for comparison. His-SUMO was used as bait in negative controls. (F) Top: effect of DPCD mutants on ciliogenesis in RPE-1 DPCD-KO cells. RPE-1 cells expressing endogenous DPCD and transfected with empty vector (EV, black) were used as control. RPE-1 DPCD-KO cells were transfected with EV (black), WT DPCD, or DPCD mutant-expressing plasmids (highlighted in red).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Hygromycin B Thermo Fisher Scientific 10687010; CAS: 31282-04-9 Tetracycline Bioshop TET701; CAS: 64-75-5 Biotin Bioshop BIO302; CAS: 58-85-5 Paraformaldehyde 32% Aqueous Solution EM Grade Electron Microscopy Sciences 15714; CAS: 30525-89-4 Bovine serum albumin Bioshop ALB001; CAS: 30525-89-4 Puromycin Bioshop PUR333; CAS: 58-58-2 JG-98 Cayman Chemicals 38337; CAS: 1456551-16-8 Geldanamycin Cayman Chemicals 13355; CAS: 30562-34-6 MK-2206 Cayman Chemicals 15569; CAS: 681281-88-9 CB-6644 Cayman Chemicals 36125; CAS: 2316817-88-4 AKTiIV (Akt Inhibitor IV) Cayman Chemicals 11593; CAS: 1032350-13-2 SIGMAFAST Protease inhibitor Tablets Sigma S8820 3x-FLAG Peptide Sigma F4799 AKT2, Active, His-tag Recombinant (phosphorylated) BPS Bioscience 40011 AKT2, Inactive, His-tag Recombinant BPS Bioscience 40001 Critical commercial assays PureLink RNA Mini Kit Thermo Fisher Scientific 12183018A QIAGEN OneStep RT-PCR Kit Qiagen 210215 Deposited data BioID mass spectrometry data This paper, deposited in MassIVE MSV000091230 DPCD SAXS model SASBDB SASDQ98 Experimental models: Cell lines HEK293 Flp-In T-REX Dr. Liliana Attisano, University of Toronto N/A HEK293T Dr. Liliana Attisano, University of Toronto N/A hTERT RPE-1 Dr. Laurence Pelletier, Lunenfeld- Tanenbaum Research Institute N/A RPE-1 DPCD KO This paper N/A RPE-1 RPAP3 KO This paper N/A RPE-1 PIH1D1 KO This paper N/A Oligonucleotides siGENOME Control Pool Non-Targeting #1 Dharmacon D-001206-13-05 siGENOME SMARTpool Human RUVBL1 Dharmacon M-008977-01-0005 siGENOME SMARTpool Human RUVBL2 Dharmacon M-012299-00-0005 siGENOME SMARTpool Human PIH1D1 Dharmacon M-020963-01-005 siGENOME SMARTpool Human DPCD Dharmacon M-027328-00-005 ON-TARGETplus SMARTpool Human RPAP3 Dharmacon L-014385-01-005 DPCD Taqman probe Thermo Fisher Scientific 4331182; assay ID: Hs00381790_m1 GAPDH Taqman probe Thermo Fisher Scientific 4448489; assay ID: Hs02786624_g1 5’-TTCAAGCTGCCACTGGC-3’ This paper DPCD sgRNA 5’-CAGCAGCTTCGGGTTCG-3’ This paper PIH1D1 sgRNA 5’-AGCACAACAGAATAAGCAGC-3’ Hart et al.77 RPAP3 sgRNA Recombinant DNA See Table S4 for list of recombinant DNA See Table S4 N/A (Continued on next page) Cell Reports 43, 113713, February 27, 2024 19

Techniques: Construct, Negative Control, Sequencing, Mutagenesis, Concentration Assay, Comparison, Expressing, Transfection, Plasmid Preparation, Control