Journal: Nature Communications

Article Title: Iontronic click-to-release enables electrically controlled delivery of drugs and biomolecules beyond charge and size limitations

doi: 10.1038/s41467-026-70985-0

Figure Lengend Snippet: a Schematic of the synthesis and immobilization strategy for biotinylated CA4-prodrug ( 6 ), synthesized via sequential decoration of cTCO-bis-NHS ( 5 ). The biotin handle allows immobilization onto streptavidin-coated magnetic beads, forming a localized, iontronically activatable prodrug reservoir. IPs were positioned above the beads in a transwell insert containing U-87 cells in the lower chamber. Grey pills indicate inactive (bound) drug, blue (colored) pills indicate active drug. b 3D-printed platform designed to precisely position iontronic devices in standard 96-well transwell plates. c Cell viability of U-87 cells after operating iontronic devices delivering 1 for 4 h at +20 nA (Active, n = 8) and at −20 nA (Reverse, n = 7) into transwells holding CA4-immobilized beads from ( a ). Points represent individual biological replicates; bars indicate mean ± SD. GC growth control (untreated, n = 32), “No Beads” = operation of iontronic devices delivering 1 at +20 nA, no beads in transwell present ( n = 6), “No Tz 1 ” = CA4-immobilized beads in transwell present with iontronic delivery of K + (0.1 M KCl) instead of Tz 1 ( n = 8). Statistical analysis was done via a Brown–Forsythe and Welch ANOVA tests (one-way) and Dunnett’s T3 multiple comparisons test. α = 0.05. p -values: GC vs. No Beads (ns) = 0.2377; GC vs. No Tz 1 (ns) = 0.3827; GC vs. Reverse, 4 h (ns) = 0.5739, GC vs. Active, 4 h (****) = <0.0001. Certain graphical elements in ( a ) were created in BioRender. Hecko, S. (2026) https://www.BioRender.com/z8yr1eu .

Article Snippet: The human GBM cell line U-87 (provided by the MUG Cell Bank, catalogue no. 300367, CLS) was cultured at 37 °C and 5% CO 2 in Eagle’s minimum essential medium (E-MEM) containing 10% fetal bovine serum (FBS), 4.5 g/L glucose, 2 mM L -glutamine, and 1% MEM Non-Essential Amino Acids.

Techniques: Synthesized, Magnetic Beads, Control

Journal: Nature Communications

Article Title: Iontronic click-to-release enables electrically controlled delivery of drugs and biomolecules beyond charge and size limitations

doi: 10.1038/s41467-026-70985-0

Figure Lengend Snippet: a Schematic overview of the bioorthogonal click-to-release (C2R) cascade of CA4-prodrug: Aminoethyl tetrazine (Tz 1 ) triggers the release of the cytotoxic agent combretastatin A-4 ( CA4 ) from sulfo-cTCO-DMEDA-CA4 ( 4 ) via a tetrazine-triggered elimination from TCO caged payload and self-immolation. b Dose-dependent cell viability of human glioblastoma cell line U-87 after 72 h incubation with the key components of the C2R reaction. Data represent mean ± SD of n = 3 independent biological replicates. Curves were fit using four-parameter logistic regression. Prodrug 4 (0.01–1000 nM, grey curve); parent CA4 (0.01–1000 nM, blue curve); released CA4 (0.01–1000 nM 4 + 5 µM 1, dashed blue curve); Tz 1 (5 µM, purple data point). c Cell viability of U-87 cells after operating iontronic devices delivering 1 for 1 h at +20 nA (Active, n = 9) and at −20 nA (Reverse, n = 11). Points represent individual biological replicates; bars indicate mean ± SD. GC growth control (untreated, n = 12). Statistical analysis was done via a Brown–Forsythe and Welch ANOVA tests (one-way) and Dunnett’s T3 multiple comparisons test. α = 0.05. p -values: GC vs. Reverse, 1 h (ns) = 0.5339; GC vs. Active, 1 h (****) = <0.0001. Certain graphical elements in ( a ) were created in BioRender. Hecko, S. (2026) https://www.BioRender.com/z8yr1eu .

Article Snippet: The human GBM cell line U-87 (provided by the MUG Cell Bank, catalogue no. 300367, CLS) was cultured at 37 °C and 5% CO 2 in Eagle’s minimum essential medium (E-MEM) containing 10% fetal bovine serum (FBS), 4.5 g/L glucose, 2 mM L -glutamine, and 1% MEM Non-Essential Amino Acids.

Techniques: Incubation, Control

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: Inhibition of collagen XVI expression reduces glioma cell invasiveness.

doi: 10.1159/000327947

Figure Lengend Snippet: Fig. 1. Analysis of COL16A1 knockdown efficiency. (A) The knockdown was translated to the protein level as shown for the full-length collagen XVI chain in cell lysates in comparison to the scr control and untreated U87MG cells. E-actin. protein expression is unaffected which indicates a specific knockdown of collagen XVI. N = 4 (B) Collagen XVI inhibited cells secreted less collagen XVI into the supernatant in comparison to scr controls. Ponceau Red stained blot membrane served as loading control. N=3 (C) Knockdown efficiency of COL16A1 was analyzed with qRT-PCR and shows an average of 60% lower COL16A1 gene expression in transfected U87MG cells (si1 and si2) compared to control cells transfected with scrambled RNA (scr). N = 19, si1= siRNA 1, si2 = siRNA 2, scr = scrambled RNA = controls.

Article Snippet: Materials and Methods Cell Culture The human glioblastoma cell line U87MG (ATCC: HTB- 14) was cultured in high glucose Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen, Paisley, UK) supplemented with 10% fetal calf serum (Sigma, St. Louis, USA) and 1% penicillin/ streptomycin (PAA, Coelbe, Germany) at 37°C in 5% CO2.

Techniques: Knockdown, Comparison, Control, Expressing, Staining, Membrane, Quantitative RT-PCR, Gene Expression, Transfection

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: Inhibition of collagen XVI expression reduces glioma cell invasiveness.

doi: 10.1159/000327947

Figure Lengend Snippet: Fig. 2. Cell morphology and number of focal adhesion contacts. (A) U87MG cells were stained using vinculin antibodies to visualize cell morphology and focal adhesion contacts. Representative images (magnification 200x and 400x) demonstrate that in collagen XVI-knockdown cells FACs were found more concentrated in the cellular periphery while in control cells FACs (white arrows) appeared to be more evenly distributed. N = 4. (B) The number of FACs per cell is significantly decreased in collagen XVI knockdown cells. The evaluation of the number of focal adhesion contacts yielded a significant decrease of 40-45% of FACs per cell after inhibition of collagen XVI. A similar reduction of focal adhesion contacts was observed for both siRNAs. N = 4; * = p < 0.05.

Article Snippet: Materials and Methods Cell Culture The human glioblastoma cell line U87MG (ATCC: HTB- 14) was cultured in high glucose Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen, Paisley, UK) supplemented with 10% fetal calf serum (Sigma, St. Louis, USA) and 1% penicillin/ streptomycin (PAA, Coelbe, Germany) at 37°C in 5% CO2.

Techniques: Staining, Knockdown, Control, Inhibition

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: Inhibition of collagen XVI expression reduces glioma cell invasiveness.

doi: 10.1159/000327947

Figure Lengend Snippet: Fig. 3. Invasive potential of collagen XVI inhibited cells. (A) Downregulation of collagen XVI expression resulted in a reduced invasion of U87MG cells in a Boyden Chamber assay compared to scr-control cells. The number of cells migrated to the lower compartment after 4 hours was significantly decreased in knockdown cells compared to the control. N = 4; * = p< 0.05; ** = p<0.01. (B, C) Adhesion capacity after 4 hours and 6 hours was not influenced by reduced collagen XVI expression. N = 6.

Article Snippet: Materials and Methods Cell Culture The human glioblastoma cell line U87MG (ATCC: HTB- 14) was cultured in high glucose Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen, Paisley, UK) supplemented with 10% fetal calf serum (Sigma, St. Louis, USA) and 1% penicillin/ streptomycin (PAA, Coelbe, Germany) at 37°C in 5% CO2.

Techniques: Expressing, Boyden Chamber Assay, Control, Knockdown

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: Inhibition of collagen XVI expression reduces glioma cell invasiveness.

doi: 10.1159/000327947

Figure Lengend Snippet: Fig. 4. Downstream gene expression analysis. (A) Gene expression of PCDH18, kindlin-1 and kindlin-2 was determined by qRT-PCR and evaluated using the ''Ct method with GAPDH as endogenous loading control set to 0 and U87MG cells transfected with scrambled RNA as calibrator. PCDH18 gene expression was profoundly upregulated whereas gene expression of kindlin-1 and kindlin-2 was clearly downregulated. N = 6. ***= p< 0.001. Protein expres- sion of PCDH 18 (B), Kindlin-1 (C) and Kindlin-2 (D) was determined with western blotting analysis of cell lysates. E-Actin was used as internal loading control. N=3. Control: cell lysate from mouse lung.

Article Snippet: Materials and Methods Cell Culture The human glioblastoma cell line U87MG (ATCC: HTB- 14) was cultured in high glucose Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen, Paisley, UK) supplemented with 10% fetal calf serum (Sigma, St. Louis, USA) and 1% penicillin/ streptomycin (PAA, Coelbe, Germany) at 37°C in 5% CO2.

Techniques: Gene Expression, Quantitative RT-PCR, Control, Transfection, Western Blot

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: Inhibition of collagen XVI expression reduces glioma cell invasiveness.

doi: 10.1159/000327947

Figure Lengend Snippet: Fig. 6. Flow cytometric analysis of E1-integrin expression and activation status. FACS analysis was performed to determine E1-integrin expression and activation on collagen XVI-knock- down U87MG cells and scr controls 96 hours post transfection. (A) The antibody integrin beta1D (MAB2259Z) recognizes activated E1-integrin only. Compared to controls E1-integrin was about 50% less activated on collagen XVI-knockdown cells. (B) The antibody integrin beta1 (MAB1900) binds to to- tal E1-integrins (both activated and inactivated forms). There- fore, the result demonstrates that total expression of E1-integrin was not altered on collagen XVI-knockdown cells. black line: Isotype control which is the secondary antibody without first antibody. si1= siRNA 1, si2 = siRNA 2, scr = scrambled RNA.

Article Snippet: Materials and Methods Cell Culture The human glioblastoma cell line U87MG (ATCC: HTB- 14) was cultured in high glucose Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen, Paisley, UK) supplemented with 10% fetal calf serum (Sigma, St. Louis, USA) and 1% penicillin/ streptomycin (PAA, Coelbe, Germany) at 37°C in 5% CO2.

Techniques: Expressing, Activation Assay, Knockdown, Transfection, Control

Journal: Scientific Reports

Article Title: Loco-regional treatment with temozolomide-loaded thermogels prevents glioblastoma recurrences in orthotopic human xenograft models

doi: 10.1038/s41598-023-31811-5

Figure Lengend Snippet: IC 50 for TMZ on cell viability in U87 MG-Red-FLuc human glioblastoma cells by cell counts. The concentration-dependent inhibitory dose-curve data was plotted as log percentage inhibition normalized to controls with applied curve non-fit calculated using GraphPad Prism. Results are shown as the mean of at least three independent experiments ± SEM.

Article Snippet: To track tumor growth in an orthotopic mouse model for glioblastoma, we decided to use commercially available human cancer cells expressing a very bright red-shifted luciferase (Red-FLuc), U87-MG-Red-FLuc cells ( https://www.perkinelmer.com/product/ivisbrite-u87mg-red-f-luc-bw124577 ).

Techniques: Concentration Assay, Inhibition

Journal: Scientific Reports

Article Title: Loco-regional treatment with temozolomide-loaded thermogels prevents glioblastoma recurrences in orthotopic human xenograft models

doi: 10.1038/s41598-023-31811-5

Figure Lengend Snippet: In vitro effect of the isolated components and of assembled new gel formulations on Glioblastoma cells. ( a ) U87-MG-Red-FLuc cells were treated with cell medium (CTRL), mesoporous SiO 2 nanoparticles (SiO 2 ) and polycaprolactone microparticles (PCL) for 24 h, causing no significant reduction in viable cells number. ( b ) U87 MG-Red-FLuc cells were treated with cell medium (CTRL), the empty thermogel (THG), and the thermogels containing mesoporous SiO 2 nanoparticles (THG@SiO 2 ) and polycaprolactone microparticles (THG@PCL) for 24 h, causing no significant reduction in viable cells number. ( c ) TMZ-containing SiO 2 nanoparticles and PCL microparticles efficiently release active drug into the cell medium to induce cell death after 24 h treatment in U87 MG-Red-FLuc cells (*p < 0.05). ( d ) THG@SiO 2 -TMZ and THG@PCL-TMZ efficiently release active TMZ into the cell medium to induce cell death after 24 h treatment in U87-MG-Red-FLuc cells (* p < 0.05; ** p < 0.005).

Article Snippet: To track tumor growth in an orthotopic mouse model for glioblastoma, we decided to use commercially available human cancer cells expressing a very bright red-shifted luciferase (Red-FLuc), U87-MG-Red-FLuc cells ( https://www.perkinelmer.com/product/ivisbrite-u87mg-red-f-luc-bw124577 ).

Techniques: In Vitro, Isolation

Journal: Scientific Reports

Article Title: Loco-regional treatment with temozolomide-loaded thermogels prevents glioblastoma recurrences in orthotopic human xenograft models

doi: 10.1038/s41598-023-31811-5

Figure Lengend Snippet: In vivo efficacy of TMZ-based therapeutic thermogels using a GBM resection and recurrence mouse model. ( a ) Experimental timeline for studying the in vivo anticancer activities of gel-based local glioblastoma treatments. Two independent experiments comparing empty THG with THG@SIO 2 -TMZ and THG@PCL-TMZ, respectively, were performed (n = 5). Results were next merged resulting in n = 8 for THG, n = 3 for THG@SIO 2 -TMZ and n = 4 for THG@PCL-TMZ, considering drop-outs due to the severity of the procedure. ( b ) Representative BLI images of mice bearing U87-MG-Red-FLuc implantation in the subcortical striatal region. After primary tumor resection, THG@SIO 2 -TMZ, THG@PCL-TMZ or empty THG were applied. ( c ) Tumor growth was directly related to the increment of luminescence signal normalized to baseline (day 2 after resection, corresponding to Time 0 in panels A and B) and recorded at days 7 (Time 1), 14 (Time 2) and 21 (Time 3) after resection, in THG, THG@SiO 2 -TMZ and THG@PCL-TMZ groups. Tumor growth was expressed as fold increase [Lum(tx)−Lum (t0)/Lum (t0)]. THG@SIO 2 -TMZ or THG@PCL-TMZ reduced tumor recurrences when compared to empty THG treatment (*p < 0.05; two-ways ANOVA p < 0.05). ( d ). Luminescence signals from control (empty THG) animals on day 21 are significantly higher (*p < 0.05; Kruskal–Wallis test p < 0.004). ( e ) Site of the resection in relation to the Paxinos representation of brain coronal section at the injection coordinate (− 0.5 mm). Representative images of H&E-stained brain sections collected 21 days after tumor resection and gel application from mice receiving either THG@SIO 2 -TMZ, THG@PCL-TMZ or empty THG (control animals) show the growth of the tumor mass to the areas adjacent to the cancer cells injection site.

Article Snippet: To track tumor growth in an orthotopic mouse model for glioblastoma, we decided to use commercially available human cancer cells expressing a very bright red-shifted luciferase (Red-FLuc), U87-MG-Red-FLuc cells ( https://www.perkinelmer.com/product/ivisbrite-u87mg-red-f-luc-bw124577 ).

Techniques: In Vivo, Injection, Staining

Journal: bioRxiv

Article Title: DEAD-box protein family member DDX28 is a negative regulator of HIF-2α and eIF4E2-directed hypoxic translation

doi: 10.1101/632331

Figure Lengend Snippet: (A) Western blot of total HIF-2α and DDX28 protein levels in normoxia (21% O 2 ) and hypoxia (1% O 2 ). GAPDH used as a loading control. (B) Western blot of DDX28 and HIF-2α (arrow, hypoxia-inducible lower band) normoxic and hypoxic protein levels in control (Ctrl) cells stably expressing a non-targeting shRNA or in cells stably expressing one of two shRNAs targeting DDX28 mRNA: Knockdown (KD) 1 and KD2. Actin used as a loading control. (C) GFP co-immunoprecipitation in hypoxic cells stably expressing FLAG-DDX28 and transfected with recombinant GFP-HIF-2α. Cells transfected with no DNA (−) or GFP alone were used as controls. (D) FLAG co-immunoprecipitation of recombinant FLAG-eIF4E2 from hypoxic cells co-transfected with HA-HIF-2α. Cells transfected with empty FLAG vector used as control. (E) FLAG co-immunoprecipitation in hypoxic cells stably expressing FLAG-DDX28 and transfected with recombinant HA-HIF-1α. Cells transfected with empty FLAG vector used as control. SE, short exposure; LE, long exposure. 25 μg of whole cell lysate was used as input. Experiments performed in U87MG glioblastoma.

Article Snippet: U87MG cells stably expressing C-terminal 3x FLAG tagged DDX28 were generated by transfecting cells with the OmicsLinkTM pEZ-M14 EX-A3144-M14 expression vector encoding the human DDX28 coding sequence (Genecopoeia).

Techniques: Western Blot, Control, Stable Transfection, Expressing, shRNA, Knockdown, Immunoprecipitation, Transfection, Recombinant, Plasmid Preparation

Journal: bioRxiv

Article Title: DEAD-box protein family member DDX28 is a negative regulator of HIF-2α and eIF4E2-directed hypoxic translation

doi: 10.1101/632331

Figure Lengend Snippet: Western blot and quantification of eIF4E2 capture with m 7 GTP-bound agarose beads in cells stably expressing one of two distinct shRNA sequences targeting DDX28 in 1% O 2 hypoxia (A-B) and 21% O 2 normoxia (C-D). 35 μg of whole cell lysate was used as the input. Ctrl, control cells stably expressing non-targeting shRNA; KD1 and KD2, knockdown cells stably expressing one of two distinct shRNA sequences targeting DDX28; LE, long exposure. Data (n ≥ 3), mean ± s.e.m normalized to input. * represents p < 0.05 using one sample t-test against hypothetical mean (μ = 1). Experiments performed in U87MG glioblastoma.

Article Snippet: U87MG cells stably expressing C-terminal 3x FLAG tagged DDX28 were generated by transfecting cells with the OmicsLinkTM pEZ-M14 EX-A3144-M14 expression vector encoding the human DDX28 coding sequence (Genecopoeia).

Techniques: Western Blot, Stable Transfection, Expressing, shRNA, Control, Knockdown

Journal: bioRxiv

Article Title: DEAD-box protein family member DDX28 is a negative regulator of HIF-2α and eIF4E2-directed hypoxic translation

doi: 10.1101/632331

Figure Lengend Snippet: Polysomal distribution of DDX28, eIF4E2 and eIF4E protein measured by western blot in control cells stably expressing non-targeting shRNA in 21% O 2 normoxia (A) and 1% O 2 hypoxia (C) and in Knockdown (KD) cells stably expressing an shRNA targeting DDX28 in normoxia (B) and hypoxia (D). Ribosomal protein L5 (rpL5) used as a marker of protein integrity in each fraction. The eIF4E (E) or eIF4E2 (F) protein associated with polysomes (fractions 4-9) as a percentage of total protein (fractions 1-9) was quantified by densitometry. Data (n = 3), mean ± s.e.m. * represents p < 0.05 using one-way ANOVA and Tukey’s HSD post-hoc test. Experiments performed in U87MG glioblastoma.

Article Snippet: U87MG cells stably expressing C-terminal 3x FLAG tagged DDX28 were generated by transfecting cells with the OmicsLinkTM pEZ-M14 EX-A3144-M14 expression vector encoding the human DDX28 coding sequence (Genecopoeia).

Techniques: Western Blot, Control, Stable Transfection, Expressing, shRNA, Knockdown, Marker

Journal: bioRxiv

Article Title: DEAD-box protein family member DDX28 is a negative regulator of HIF-2α and eIF4E2-directed hypoxic translation

doi: 10.1101/632331

Figure Lengend Snippet: ( A) Western blot of HIF-2α protein levels in cytoplasmic and nuclear fractions of control cells expressing a non-targeting shRNA or Knockdown (KD) cells stably expressing an shRNA targeting DDX28 in 1% O 2 hypoxia. Lamin a/c used as nuclear marker and α-tubulin as cytoplasmic marker. (B) The mRNA abundance of HIF-2α gene targets in hypoxia measured via qRT-PCR. Data (n ≥ 3), mean ± s.e.m. represented as log 2 (fold change) in DDX28 KD cells relative to control cells and normalized to endogenous control genes RPLP0 and RPL13A. * represents p < 0.05 using a one-sample t-test against hypothetical mean (μ = 0). CITED2, Cbp/P300-Interacting Transactivator 2; EPO, Erythropoietin; IGFBP3, Insulin Like Growth Factor Binding Protein 3; ITPR1, inositol 1,4,5-trisphosphate receptor type 1; LOXL2, Lysyl Oxidase Like 2; OCT4, octamer-binding transcription factor 4. Experiments performed in U87MG glioblastoma.

Article Snippet: U87MG cells stably expressing C-terminal 3x FLAG tagged DDX28 were generated by transfecting cells with the OmicsLinkTM pEZ-M14 EX-A3144-M14 expression vector encoding the human DDX28 coding sequence (Genecopoeia).

Techniques: Western Blot, Control, Expressing, shRNA, Knockdown, Stable Transfection, Marker, Quantitative RT-PCR, Binding Assay

Journal: bioRxiv

Article Title: DEAD-box protein family member DDX28 is a negative regulator of HIF-2α and eIF4E2-directed hypoxic translation

doi: 10.1101/632331

Figure Lengend Snippet: Viable cell counts were measured with crystal violet staining after 24 h, 48 h, and 72 h in 21% O 2 normoxia (A) and 1% O 2 hypoxia (B) for control cells expressing a non-targeting shRNA or Knockdown cells stably expressing one of two shRNAs targeting DDX28 (KD1 and KD2). All absolute cell count values were normalized to the number of cells present on day 0 for each individual independent experiment, representing the fold change in the number of cells at each time point relative to day 0. Proliferation was measured as % BrdU-positive control and DDX28 KD cells via immunofluorescence after 24 h in normoxia (C) or hypoxia (D). Data (n ≥ 3), mean ± s.e.m. * represents p < 0.05 using an unpaired two-sample t-test. Experiments performed in U87MG glioblastoma.

Article Snippet: U87MG cells stably expressing C-terminal 3x FLAG tagged DDX28 were generated by transfecting cells with the OmicsLinkTM pEZ-M14 EX-A3144-M14 expression vector encoding the human DDX28 coding sequence (Genecopoeia).

Techniques: Staining, Control, Expressing, shRNA, Knockdown, Stable Transfection, Cell Counting, Positive Control, Immunofluorescence