human hcc44 cell line  (DSMZ)

Journal: The EMBO Journal

Article Title: Ubiquitin pathway blockade reveals endogenous ADP-ribosylation marking PARP7 and AHR for degradation

doi: 10.1038/s44318-025-00656-1

Figure Lengend Snippet: ( A ) Schematic overview of the ubiquitin-proteasome system, major PARPs and their inhibitors used in this study. Ub: ubiquitin, E1: ubiquitin-activating enzyme, E2: ubiquitin conjugating enzyme, E3: ubiquitin ligase (HECT or RING type). ( B ) HCC44 cells were treated with DMSO or 1 μM TAK243 for 4 h. ( C ) Time-course experiment with TAK243 and MG132 at 1 and 10 μM, respectively, in HCC44 cells. ADPr was assessed by western blotting using the anti-poly/mono-ADPr antibody, with α-tubulin as a loading control. .

Article Snippet: Human HCC44 cell line was purchased from the German Collection of Microorganisms and Cell Cultures (#ACC 534, DSMZ, Braunschweig, Germany).

Techniques: Ubiquitin Proteomics, Western Blot, Control

Journal: The EMBO Journal

Article Title: Ubiquitin pathway blockade reveals endogenous ADP-ribosylation marking PARP7 and AHR for degradation

doi: 10.1038/s44318-025-00656-1

Figure Lengend Snippet: ( A ) HCC44 cells pre-treated with DMSO or PARP inhibitors for 20 h were further treated with 1 µM TAK243 for 4 h. ADPr was assessed by western blotting using the anti-poly/mono-ADPr antibody, with α-tubulin as a loading control. ( B ) HiBiT-PARP7 CT26 cells pre-treated with DMSO or PARP inhibitors for 20 h were further treated with 1 µM TAK243 for 4 h. ADPr was assessed by western blotting using the anti-poly/mono-ADPr antibody, with α-tubulin as a loading control. Overlap of the signals was obtained using fluorescently labelled anti-rabbit (poly/mono-ADPr, green) and anti-mouse (HiBiT, red) secondary antibodies. ( C ) HiBiT luminescence assay in HiBiT-PARP7 CT26 cells with 1 µM TAK243, 10 µM MG132, 100 nM RBN2397 (PARP7i) treatments for 4 h showing induction of PARP7 levels (HiBiT luminescence). Wild-type CT26 cells (non-HiBiT) were used as a negative control. Data are shown as mean ± s.e.m. of n = 3 technical replicates. ( D ) A549 cells were co-treated with PARP inhibitors and 1 µM TAK243 for 24 h. PARP inhibitor concentrations in panels (A– C ) are specified in the methods section. ADPr was assessed by western blotting using the anti-mono-ADPr antibody. α-tubulin was used as a loading control. The following PARP inhibitors were used: PARP1/2 (1 µM olaparib), PARP7 (100 nM RBN2397), PARP14 (RBN012759, 200 nM in Fig. 2A, B and 500 nM in Fig. 2D), TNKS (1 µM AZ6102). PARP14, PARP7, TNKS and PARP1 were detected with corresponding antibodies in relevant panels. .

Article Snippet: Human HCC44 cell line was purchased from the German Collection of Microorganisms and Cell Cultures (#ACC 534, DSMZ, Braunschweig, Germany).

Techniques: Western Blot, Control, Luminescence Assay, Negative Control

Journal: The EMBO Journal

Article Title: Ubiquitin pathway blockade reveals endogenous ADP-ribosylation marking PARP7 and AHR for degradation

doi: 10.1038/s44318-025-00656-1

Figure Lengend Snippet: ( A ) Schematic representation of AHR activation and degradation. Tapinarof is an agonist of AHR. ( B ) HCC44 cells were pre-treated with DMSO, 1 µM tapinarof and PARP inhibitors (PARP1/2 (1 µM olaparib), PARP7 (100 nM RBN2397), PARP14 (200 nM RBN012759), TNKS (1 µM AZ6102)) for 20 h followed by an additional 4 h of 1 µM TAK243 or 10 µM MG132 treatment. ( C ) MCF7 wild type (WT) and PARP7 knockout (KO) cells were treated as in ( B ). ADPr was assessed by western blotting using the anti-poly/mono-ADPr antibody. AHR, PARP7, TNKS and PARP14 were detected using corresponding antibodies with α-tubulin as a loading control. .

Article Snippet: Human HCC44 cell line was purchased from the German Collection of Microorganisms and Cell Cultures (#ACC 534, DSMZ, Braunschweig, Germany).

Techniques: Activation Assay, Knock-Out, Western Blot, Control

Journal: The EMBO Journal

Article Title: Ubiquitin pathway blockade reveals endogenous ADP-ribosylation marking PARP7 and AHR for degradation

doi: 10.1038/s44318-025-00656-1

Figure Lengend Snippet: ( A ) HCC44 cells were transfected with siRNAs against DELTEX E3 ligases and other ADP-ribose-binding E3 ligases, treated with DMSO or 1 µM tapinarof for 24 h, and analysed by western blotting. ( B ) HCC44 cells transfected with two separate DTX2 siRNAs, treated with DMSO or 1 µM tapinarof for 24 h, and analysed by western blotting. ( C ) A549 cells transfected with two separate DTX2 siRNAs, treated with DMSO or 1 µM tapinarof for 24 h, and analysed by western blotting. ADPr was assessed with a poly-mono-ADPr antibody, DTX2 knockdown was confirmed with an anti-DTX2 antibody, α-tubulin was used as a loading control. Overlap of the signals was obtained using fluorescently labelled anti-rabbit (poly/mono-ADPr, green) and anti-mouse (AHR, red) secondary antibodies. .

Article Snippet: Human HCC44 cell line was purchased from the German Collection of Microorganisms and Cell Cultures (#ACC 534, DSMZ, Braunschweig, Germany).

Techniques: Transfection, Binding Assay, Western Blot, Knockdown, Control

Journal: The EMBO Journal

Article Title: Ubiquitin pathway blockade reveals endogenous ADP-ribosylation marking PARP7 and AHR for degradation

doi: 10.1038/s44318-025-00656-1

Figure Lengend Snippet: ( A ) Widefield images showing HCC44 cells treated with 1 µM tapinarof (24 h), 1 µM TAK243 (4 h) and 100 nM PARP7i (RBN2397, 24 h) alone and in combination as in Fig. . Cells were stained with Hoechst (blue), ADPr (Poly/mono-ADP-ribose, green) and AHR (magenta). ( B , C ) Quantification of nuclear ADPr ( B ) or AHR ( C ) from ( A ). Between 5156 and 6220 cells were measured in each condition. ( D ) Widefield images showing A549 WT or DTX2 knockout (KO) cells treated with 1 μM tapinarof (24 h), 1 μM TAK243 (4 h) and 100 nM PARP7i (RBN2397, 24 h) alone and in combination as in Fig. . Cells were stained with Hoechst (blue), ADPr (poly/mono-ADP-ribose, green) and AHR (magenta). ( E , F ) Quantification of nuclear ADPr ( E ) or AHR ( F ) from ( D ). Between 5716 and 13282 cells were measured in each condition. ( G ) Widefield images showing CT26 (HiBiT-PARP7) cells treated with 1 μM TAK243 (4 h) and 100 nM PARP7i (RBN2397, 24 h) alone and in combination, as in Fig. . Cells were stained with Hoechst (blue), ADPr (poly/mono-ADP-ribose, green) and HiBiT (magenta). ( H , I ) Quantification of nuclear ADPr ( B ) or HiBiT ( I ) from ( G ). Between 2991 and 6701 cells were measured in each condition. For all images, scale bar = 20 μm. Statistical analysis was performed using an ordinary one-way ANOVA. Asterisks indicate statistical significance (**** P < 0.0001). Red bars indicate the median for each condition. .

Article Snippet: Human HCC44 cell line was purchased from the German Collection of Microorganisms and Cell Cultures (#ACC 534, DSMZ, Braunschweig, Germany).

Techniques: Staining, Knock-Out

Journal: Scientific reports

Article Title: Thyroid Transcription Factor 1 Reprograms Angiogenic Activities of Secretome.

doi: 10.1038/srep19857

Figure Lengend Snippet: Figure 2. TTF-1 upregulates VEGF. Three dox-on TTF-1-inducible cells systems were hosted in two human lung AD cell lines (NCI-H1792 and HCC44) and a premalignant human lung epithelial cell line (BEAS-2B). A dox-on transgene of the HDD mutant of TTF-1 was also created in HCC44 and BEAS-2B cells. (A) TTF- 1− human lung AD cell line (A549) was used to host stable expression of either wt-TTF-1 or HDD mutant transgene. Protein expression of the dox-on TTF-1 transgene is shown in A, whereas the VEGF RNA (B,D,F,H) of the transfectant cells and the secreted VEGF protein in the corresponding CM (C,E,G,I) demonstrate VEGF upregulation by TTF-1. EV, empty vector.

Article Snippet: Human lung cancer cell line HCC44 was obtained from the Leibniz Institute (DSMZ) and maintained in RPMI-1640 supplemented with 10% fetal bovine serum (FBS), penicillin and streptomycin.

Techniques: Mutagenesis, Expressing, Transfection, Plasmid Preparation