Journal: Cancers

Article Title: Piezo1-ATF3-PPP1r15a Axis Transduces Mechanical Stress into Apoptosis in Glioma Under Low-Intensity Focused Ultrasound

doi: 10.3390/cancers18091445

Figure Lengend Snippet: ATF3 is involved in the ERS of GBM cells treated with LIFU. ( A ) Annexin-V/PI bound U251 or LN229 cells were counted. ( B ) The corresponding statistical results of apoptosis rate. ( C ) The expression levels of Bax, Bcl-2 and caspase3 were determined by Western blot. ( D ) The expression levels of ATF3, ATF4 and CHOP were determined by Western blot. ( E ) U251 and LN229 cells were subjected to reverse transcription-quantitative PCR analysis for ATF4. ( F ) U251 and LN229 cells were subjected to reverse transcription-quantitative PCR analysis for CHOP. ( G ) Representative images of JC-1 staining in U251 with different treatments. ( H ) Representative images of JC-1 staining in LN229 with different treatments. The data are presented as the means ± SEM from at least three independent biological replicates. Statistical comparisons among multiple groups (control, LIFU, siATF3, LIFU + siATF3) were performed using two-way ANOVA followed by Šídák’s multiple comparisons test. *** p < 0.001; **** p < 0.0001. ER, endoplasmic reticulum; GBM, glioblastoma; LIFU, low-intensity focused ultrasound. The original Western blot figures can be found in .

Article Snippet: The American Type Culture Collection provided GBM cell lines (U251 and LN229) and the 293T cell line.

Techniques: Expressing, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Staining, 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 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: Cells

Article Title: Serglycin Cooperates with the Unfolded Protein Response Pathway and Inflammation to Drive Glioblastoma Cell Survival

doi: 10.3390/cells15080660

Figure Lengend Snippet: Loss of SRGN diminishes the expression of UPR markers in GBM cell lines and results in ER stress susceptibility. ( A ) Assessment of the mRNA levels of UPR mediators in LN-18 shSCR and LN-18 shSRGN cells through real-time qPCR. ( B ) Relative mRNA levels of SRGN in LN-18 and U251-MG cell lines. ( C ) SRGN suppression levels in U251-MG siSRGN cells via real-time qPCR and ( D ) phase-contrast microscopy images upon SRGN silencing in U251-MG cells (48 h). ( E ) Relative proliferation rates of U251-MG siControl and U251-MG siSRGN cells after transient SRGN silencing (48 h). ( F ) Evaluation of the gene expression of UPR markers in U251-MG siControl and U251-MG siSRGN cells (48 h). ( G ) IC50 values for LN-18 cell lines upon TM treatment (48 h). ( H ) Wound closure assay in LN-18 cell lines after TM treatment for 24 h and 48 h. Statistically significant differences compared to control are shown by asterisk: * ( p ≤ 0.05).

Article Snippet: The LN-18 GBM cell line was obtained from the American Type Culture Collection (ATCC).

Techniques: Expressing, Microscopy, Gene Expression, Wound Closure Assay, Control

Journal: Cells

Article Title: Serglycin Cooperates with the Unfolded Protein Response Pathway and Inflammation to Drive Glioblastoma Cell Survival

doi: 10.3390/cells15080660

Figure Lengend Snippet: SRGN depletion attenuates the activation profile of UPR in GBM cell lines and renders cells non-responsive to ER stress. ( A ) Western blot analysis of UPR effectors’ protein and phosphorylated levels in TM-treated (48 h) LN-18 cells. ( B ) Immunofluorescence staining for ATF4 (green) and nuclei (blue) depicting the distribution and nuclear accumulation of ATF4 in TM-treated (48 h) LN-18 shSCR cells. ATF4 was not detected in LN-18 shSRGN cells under relative exposure. Scale bar 25 μm. ( C ) Western blot analysis of the basal UPR protein and phosphorylated levels in U251-MG siControl and U251-MG siSRGN cells, following transient SRGN silencing (72 h). All blots are representative of at least three independent experimental repetitions.

Article Snippet: The LN-18 GBM cell line was obtained from the American Type Culture Collection (ATCC).

Techniques: Activation Assay, Western Blot, Immunofluorescence, Staining

Journal: Cells

Article Title: Serglycin Cooperates with the Unfolded Protein Response Pathway and Inflammation to Drive Glioblastoma Cell Survival

doi: 10.3390/cells15080660

Figure Lengend Snippet: SRGN-expressing cells possess a pro-survival UPR mechanism and demand global UPR activation. Assessment of LN-18 shSCR and LN-18 shSRGN cells’ viability upon treatment with ( A ) MKC8866 (IRE1 RNase activity inhibitor), ( B ) Kira6 (IRE1 kinase activity inhibitor), ( C ) GSK2606414 (PERK kinase activity inhibitor) and ( D ) Ceapin-A7 (ATF6 TF activity inhibitor) for 24 h incubation at 0.1, 1 and 10 μΜ final concentrations. Statistically significant differences compared to control are shown by bars and asterisk: * ( p ≤ 0.05).

Article Snippet: The LN-18 GBM cell line was obtained from the American Type Culture Collection (ATCC).

Techniques: Expressing, Activation Assay, Activity Assay, Incubation, Control

Journal: Cells

Article Title: Serglycin Cooperates with the Unfolded Protein Response Pathway and Inflammation to Drive Glioblastoma Cell Survival

doi: 10.3390/cells15080660

Figure Lengend Snippet: SRGN suppression provokes an apoptotic phenotype in GBM cells. ( A ) Western blot analysis of the intact and cleaved forms of caspase-3 and PARP-1, as well as the active and total Akt forms in TM-treated (48 h) LN-18 cells. ( B ) caspase-3 and PARP-1 proteolysis assessed through Western blotting in U251-MG cells after transient SRGN knock-down (72 h). ( C ) Zymography assay for determining the active intracellular CTSB isoforms in acidic pH upon TM treatment (48 h) in LN-18 cell lines. Real-time qPCR for evaluating the relative mRNA levels of ( D ) CTSB , ( E ) BAX , ( F ) Bcl-2 and ( G ) Beclin-1 in TM-treated (48 h) LN-18 cell lines. ( H ) Basal mRNA levels of BAX , Bcl-2 and Beclin-1 in U251-MG cells upon SRGN depletion (48 h). All blots and zymography images are representative of at least three independent experimental repetitions. Statistically significant differences compared to control are shown by bars and asterisk: * ( p ≤ 0.05).

Article Snippet: The LN-18 GBM cell line was obtained from the American Type Culture Collection (ATCC).

Techniques: Western Blot, Knockdown, Zymography, Control

Journal: Cells

Article Title: Serglycin Cooperates with the Unfolded Protein Response Pathway and Inflammation to Drive Glioblastoma Cell Survival

doi: 10.3390/cells15080660

Figure Lengend Snippet: SRGN orchestrates the inflammatory response in GBM cells. ( A ) Relative mRNA levels of TLRs and TNFRs in LN-18 shSCR and LN-18 shSRGN cell lines assessed through real-time PCR. ( B ) Immunofluorescence staining for P-p65 (green) and nuclei (blue) depicting the distribution and nuclear accumulation of phosphorylated p65 subunit in LN-18 cells at basal conditions. P-p65 was not detected in LN-18 shSRGN cells under relative exposure. Scale bar 25 μm. Real-time qPCR for evaluating the relative mRNA levels of ( C ) TLR2 , ( D ) TLR4 , ( E ) TNFRI , ( F ) TNFRII , ( G ) IL-1β , ( H ) IL-8 and ( I ) CXCL-1 in TM-treated (48 h) LN-18 cell lines. ( J ) Basal mRNA levels of TLR4 and IL-1β in U251-MG cells upon transient SRGN silencing (48 h) assessed through real-time qPCR. ( K ) p65/NF-kB phosphorylated and total forms in TM-treated (48 h) LN-18 cell lines evaluated by Western blotting. All blots are representative of at least three independent experimental repetitions. Statistically significant differences compared to control are shown by bars and asterisk: * ( p ≤ 0.05).

Article Snippet: The LN-18 GBM cell line was obtained from the American Type Culture Collection (ATCC).

Techniques: Real-time Polymerase Chain Reaction, Immunofluorescence, Staining, Western Blot, Control