Journal: Oncotarget

Article Title: ErbB2 inhibition by lapatinib promotes degradation of mutant p53 protein in cancer cells

doi: 10.18632/oncotarget.12878

Figure Lengend Snippet: ( A ) Lapatinib inhibits HSF1Ser326 phosphorylation induced by heat shock in BT474 cells. Cells were pre-treated with 300nM of lapatinib for 24 h. After heat shock (42°C, 30 min) cells were immediately analyzed by immunoblot. ( B ) Heat shock (42°C, 30 min) alleviates mutp53 destabilization by lapatinib in p53Q/-;ErbB2 MECs. Cells were pre-treated with lapatinib (300 nM, 24 h). After heat shock (42°C, 30 min) cells were immediately analyzed by immunoblot. ( C ) Lapatinib (300 nM) mediated mutp53 destabilization coincides with reduction of HSF1 levels in Q/-;ErbB2, but not in p53−/−;ErbB2 MECs. ( D ) In HSF1-ablated BT474 cells, lapatinib (300 nM) does not induce further destabilization of mutp53. Cells were transfected with siHSF1 or scrambled siRNA control. After 24 h, cells were treated with 300 nM of lapatinib for an additional 24 h followed by immunoblot. ( E ) Lapatinib destabilizes mutp53 only in the presence of functional Hsp90. BT474cells were pre-treated with ganetespib (250 nM) for 6 h followed by lapatinib (250 nM and 500 nM) treatment for 24 h. Lapatinib even at high concentrations does not further decrease mutp53 and MDM2 levels in cells pre-treated with Hsp90 inhibitor.

Article Snippet: For immunoblots, equal total protein of cell lysates (2.5–20 μg) were detected with antibodies to mouse p53 (FL393), human p53 (PAb1801), MDM2, GAPDH, HSC 70 (all Santa Cruz Biotechnology), Erk, pErk (T202/Y204), EGFR, EGRF-Tyr845P, ErbB2, pErbB2 (Y1221/1222) (all Cell Signaling), HSF1, pSer326 HSF1, Hsp90 (Enzo Life Sciences, Inc., Farmingdale, NY).

Techniques: Phospho-proteomics, Western Blot, Transfection, Control, Functional Assay

Journal: Oncotarget

Article Title: ErbB2 inhibition by lapatinib promotes degradation of mutant p53 protein in cancer cells

doi: 10.18632/oncotarget.12878

Figure Lengend Snippet: ( A ) Lapatinib shows preferential cytotoxicity in mutp53;ErbB2 mammary cells. H/H;ErbB2 MECs show higher sensitivity to lapatinib, compared to p53−/−;ErbB2 counterparts in colony formation assay (crystal violet staining). H/H;ErbB2 and −/−;ErbB2 MECs were treated with 1 μM of lapatinib for 5 weeks. ( B ) H/H;ErbB2 MECs show higher viability loss in response to lapatinib compered to −/−;ErbB2 and H/H MECs. Cells were treated with indicated concentrations of lapatinib for 48h. Viability loss is shown relative to DMSO treated controls. ( C ) Lapatinib destabilizes mutp53 but not wtp53. Lapatinib does not affect wild-type p53 protein even at high concentrations. MCF7 cells were treated with indicated concentrations of lapatinib for 24 h. ( D ) Mammary tumor cell lines show both elevated ErbB2 and pErbB2 levels (Y1221/1222) in the presence of the mutp53 allele compared with p53 +/+;ErbB2. Cells were established from primary mammary tumors of mice with indicated genotypes. ( E ) Heat shock (42°C, 30 min) more potently induces Ser326HSF1 activation in the presence of mutp53 allele compared with p53+/+;ErbB2 tumor cell lines. After heat shock p53+/+;ErbB2 (#1761) and H/+;ErbB2 (#1349) cells were immediately analyzed by immunoblot. ( F ) Lapatinib more profoundly inhibits of ErbB2 downstream signaling (pErk, pErbB2, HSF1) in H/+;ErbB2 compared with p53+/+;ErbB2 cancer cells. Cells were treated with lapatinib (0.6 μM, 24 h) and analyzed by immunoblot. ( G ) Tumor cell lines harboring mutp53 allele respond better to lapatinib measured by cell viability assay. Mammary tumor cell lines with indicated genotypes were treated with 0.5 and 1 μM of lapatinib for 48 h. Viability loss is shown relative to DMSO treated controls (calculated as 100% viability).

Article Snippet: For immunoblots, equal total protein of cell lysates (2.5–20 μg) were detected with antibodies to mouse p53 (FL393), human p53 (PAb1801), MDM2, GAPDH, HSC 70 (all Santa Cruz Biotechnology), Erk, pErk (T202/Y204), EGFR, EGRF-Tyr845P, ErbB2, pErbB2 (Y1221/1222) (all Cell Signaling), HSF1, pSer326 HSF1, Hsp90 (Enzo Life Sciences, Inc., Farmingdale, NY).

Techniques: Colony Assay, Staining, Activation Assay, Western Blot, Viability Assay

Journal: Oncotarget

Article Title: ErbB2 inhibition by lapatinib promotes degradation of mutant p53 protein in cancer cells

doi: 10.18632/oncotarget.12878

Figure Lengend Snippet: ErbB2 signaling mediates HSF1 activation in a mutp53-dependent manner. Mutp53, by enhancing ErbB2 signaling, potentiates HSF1 activity via a feed-forward loop and thereby upregulates Hsp90 clients, including mutp53. Inhibition of ErbB2 by lapatinib, leads to inhibition of HSF1 transcriptional function, decline in Hsp90 level, release MDM2 from inhibitory complex and subsequent degradation of mutp53 and MDM2.

Article Snippet: For immunoblots, equal total protein of cell lysates (2.5–20 μg) were detected with antibodies to mouse p53 (FL393), human p53 (PAb1801), MDM2, GAPDH, HSC 70 (all Santa Cruz Biotechnology), Erk, pErk (T202/Y204), EGFR, EGRF-Tyr845P, ErbB2, pErbB2 (Y1221/1222) (all Cell Signaling), HSF1, pSer326 HSF1, Hsp90 (Enzo Life Sciences, Inc., Farmingdale, NY).

Techniques: Activation Assay, Activity Assay, Inhibition

Journal: Frontiers in Microbiology

Article Title: Postbiotics derived from recombinant lactic acid bacteria exhibit high IL6-binding capacity and suppress IL6-induced STAT3 signaling

doi: 10.3389/fmicb.2025.1657810

Figure Lengend Snippet: Killing efficiency of different physicochemical treatments on recombinant L. lactis strains displaying different binders of proinflammatory cytokines on their surface. The killing efficiency was determined by culturing and counting colonies on agar plates. Complete killing of bacteria i.e., 0 CFU/mL (no bacterial colonies on agar plates) was considered as 100% efficiency. pSD-ZIL6, L. lactis displaying anti-IL6 affibody; pSD-ZTNF, L. lactis displaying anti-TNF affibody; pSD-Fyn17, L. lactis displaying anti-IL17 fynomer; pSD-EVA, L. lactis displaying anti-IL8 evasin; pNZ8148, L. lactis harboring empty plasmid. The results presented are means ± standard deviation (SD) of three technical replicates from a representative experiment.

Article Snippet: Briefly, serial dilutions of IL6 (0.32, 0.65, 1.31, 2.62, 5.25, 10.5, 21, 42 nM; HZ-1019, Proteintech) were incubated with a fixed number of recombinant bacteria (8 × 10 6 CFU) for 2 h in a final volume of 200 μl.

Techniques: Recombinant, Bacteria, Plasmid Preparation, Standard Deviation

Journal: Frontiers in Microbiology

Article Title: Postbiotics derived from recombinant lactic acid bacteria exhibit high IL6-binding capacity and suppress IL6-induced STAT3 signaling

doi: 10.3389/fmicb.2025.1657810

Figure Lengend Snippet: Cytokine binding ability of recombinant L. lactis bacteria displaying different binders of proinflammatory cytokines on their surface upon exposure to bacteria-killing treatments. ELISA-determined concentrations of recombinant IL6 (A) , TNF (B) , IL17 (C) , and IL8 (D) that remained in the solution following incubation with the corresponding strain of bacteria before and after treatment with heat (70 °C, 40 min or 100 °C, 30 min), sonication, ethanol, UV, and gamma irradiation. pSD-ZIL6, L. lactis displaying ZIL6 affibody; pSD-ZTNF, L. lactis displaying anti-TNF affibody; pSD-Fyn17, L. lactis displaying anti-IL17 fynomer; pSD-EVA, L. lactis displaying anti-IL8 evasin; Ctrl: L. lactis control cells containing empty plasmid pNZ8148. The results are presented as means ± SD of three technical replicates of a representative experiment. ns, p = 0.09; *, p ≤ 0.05; **, p = 0.002; ***, p < 0.001 (unpaired two-tailed t -test).

Article Snippet: Briefly, serial dilutions of IL6 (0.32, 0.65, 1.31, 2.62, 5.25, 10.5, 21, 42 nM; HZ-1019, Proteintech) were incubated with a fixed number of recombinant bacteria (8 × 10 6 CFU) for 2 h in a final volume of 200 μl.

Techniques: Binding Assay, Recombinant, Bacteria, Enzyme-linked Immunosorbent Assay, Incubation, Sonication, Irradiation, Control, Plasmid Preparation, Two Tailed Test

Journal: Frontiers in Microbiology

Article Title: Postbiotics derived from recombinant lactic acid bacteria exhibit high IL6-binding capacity and suppress IL6-induced STAT3 signaling

doi: 10.3389/fmicb.2025.1657810

Figure Lengend Snippet: Determination of binding affinity of ZIL6-displaying L. lactis for human IL6. A constant number of 8 × 10 6 CFU bacteria was incubated with increasing concentrations of human IL6 for 2 h. MFI values were measured by flow cytometry and binding curves were fitted to the data using the Hill equation in the GraphPad Prism v.10.3.1. The dissociation constants (Kd) values were calculated from the binding curves for non-treated, live ZIL6-displaying L. lactis bacteria and bacteria treated by heat (70 °C, 40 min or 100 °C, 30 min), sonication, ethanol, and UV irradiation. Data are means ± SD of three technical replicates from a representative experiment.

Article Snippet: Briefly, serial dilutions of IL6 (0.32, 0.65, 1.31, 2.62, 5.25, 10.5, 21, 42 nM; HZ-1019, Proteintech) were incubated with a fixed number of recombinant bacteria (8 × 10 6 CFU) for 2 h in a final volume of 200 μl.

Techniques: Binding Assay, Bacteria, Incubation, Flow Cytometry, Sonication, Irradiation

Journal: Frontiers in Microbiology

Article Title: Postbiotics derived from recombinant lactic acid bacteria exhibit high IL6-binding capacity and suppress IL6-induced STAT3 signaling

doi: 10.3389/fmicb.2025.1657810

Figure Lengend Snippet: Quantification of the maximum binding capacity of ZIL6-displaying L. lactis . Increasing concentrations of recombinant human IL6 (0.45, 4.5, 45 and 450 ng) were incubated with a constant number of bacteria (4.5 × 10 7 CFU equivalent to 0.1 mg dry cell weight) in 450 μl. Residual IL6 in the supernatants was quantified by ELISA for live non-treated bacteria and the cells treated by heat (70 °C, 40 min or 100 °C, 30 min), sonication, ethanol, and UV irradiation. The percentage of bound IL6 was calculated from the difference measured in the presence of ZIL6-displaying L. lactis and control bacteria (carrying empty plasmid pNZ8148). The data are means ± SD of two biological replicates. The asterisks denote statistically significant differences between treated and non-treated bacteria for each concentration. ***, p < 0.001 (one-way ANOVA with Dunnett multiple comparison test).

Article Snippet: Briefly, serial dilutions of IL6 (0.32, 0.65, 1.31, 2.62, 5.25, 10.5, 21, 42 nM; HZ-1019, Proteintech) were incubated with a fixed number of recombinant bacteria (8 × 10 6 CFU) for 2 h in a final volume of 200 μl.

Techniques: Binding Assay, Recombinant, Incubation, Bacteria, Enzyme-linked Immunosorbent Assay, Sonication, Irradiation, Control, Plasmid Preparation, Concentration Assay, Comparison

Journal: Frontiers in Microbiology

Article Title: Postbiotics derived from recombinant lactic acid bacteria exhibit high IL6-binding capacity and suppress IL6-induced STAT3 signaling

doi: 10.3389/fmicb.2025.1657810

Figure Lengend Snippet: (A) Inhibition of STAT3 signaling in HEKblue-IL6R cells by non-viable ZIL6-displaying L. lactis bacteria in comparison to live non-treated strain. ZIL6-displaying L. lactis cells (1 × 10 8 CFU/mL or 1 × 10 9 CFU/mL) were preincubated with IL6 (1 ng/mL) for 2 h. After removal of bacterial cells, the cell-free supernatant containing remainder of IL6 was added to HEKblue-IL6R cells to induce the reporter system. The percentage of STAT3 inhibition was calculated relative to the STAT3 signaling induced by IL6 in the absence of bacteria. The inhibition of STAT3 signaling was determined for live non-treated bacteria and the cells treated by heat (70 °C, 40 min or 100 °C, 30 min), sonication, ethanol, UV, and gamma irradiation. pSD-ZIL6, L. lactis displaying ZIL6 affibody; Ctrl: L. lactis control cells containing empty plasmid pNZ8148. An anti-IL6 monoclonal antibody (Ab) was used as a positive control. The data are means ± SD of four biological replicates. The asterisks denote statistically significant differences between treated and non-treated bacteria for each concentration. ***, p < 0.001 (one-way ANOVA with Dunnett multiple comparison test). (B) The effect of lactic acid production by L. lactis on viability of HEKblue-IL6R cells. HEKblue-IL6R cells (100 000 cells/well) were incubated with ZIL6-displaying L. lactis (2 × 10 7 bacteria/well) for 6, 12, and 24 h, and the viability of HEKblue-IL6R cells was determined with trypan blue. The data are means ± SD of three technical replicates of a representative experiment.

Article Snippet: Briefly, serial dilutions of IL6 (0.32, 0.65, 1.31, 2.62, 5.25, 10.5, 21, 42 nM; HZ-1019, Proteintech) were incubated with a fixed number of recombinant bacteria (8 × 10 6 CFU) for 2 h in a final volume of 200 μl.

Techniques: Inhibition, Bacteria, Comparison, Sonication, Irradiation, Control, Plasmid Preparation, Positive Control, Concentration Assay, Incubation

Journal: Organic & biomolecular chemistry

Article Title: Novel aryltriazole acyclic C-azanucleosides as anticancer candidates.

doi: 10.1039/d0ob02164d

Figure Lengend Snippet: Fig. 4 Compound If efficiently inhibited the expression of HSF1, HSP27, HSP70, HSP90α and eIF4E in Panc-1 (A) and HepG2 cells (B). Panc-1 and HepG2 cells were treated with If at indicated concentrations. Protein levels of HSF1, HSP27, HSP70, HSP90α and eIF4E were then analyzed by western blotting, with β-actin as the reference.

Article Snippet: The primary antibodies including PARP antibody, BCL-2 antibody, HSF1 rabbit mAb, HSP70 rat mAb, HSP27 rabbit mAb and eIF4E rabbit mAb were all obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Western Blot

Journal: ACS applied bio materials

Article Title: Attenuation of Abnormal Scarring Using Spherical Nucleic Acids Targeting Transforming Growth Factor Beta 1

doi: 10.1021/acsabm.0c00990

Figure Lengend Snippet: Cellular uptake of SNAs and TGF-β1 regulation in fibroblasts. (A) Cellular uptake of Cy3-labeled AuSNAs in Rab9 cells visualized with confocal microscopy. Fluorescence from SNAs is shown in red and blue is a nuclear stain (DAPI). Scale bar = 10 μm. Downregulation of TGF-β1 by TGF-β1 AuSNAs and LSNAs in (B) Rab9 cells, (C) HSF cells, and (D) KF cells. NS = nonsense.

Article Snippet: Rab9 cells were purchased from American Type Culture Collection (CRL-1414) and HSF cells were purchased from CellResearch Corporation (HSF107).

Techniques: Labeling, Confocal Microscopy, Fluorescence, Staining

Journal: Scientific Reports

Article Title: Quercetin can reduce viral RNA level of O’nyong-nyong virus and resulting innate immune cytokine responses in cultured human synovial fibroblasts

doi: 10.1038/s41598-021-85840-z

Figure Lengend Snippet: Concentration-dependent cytotoxic effects of phenolics or ONNV on cultured HSF cells. Cells were infected by ONNV (MOI 10 –3 to 1) or phenolics (1 to 100 µM) for 24 h. Released LDH in culture medium was measured using a colorimetric-based method (CytoTox 96 Non-Radioactive Cytotoxicity Assay) and expressed relative to the maximum release by application of a lysis buffer (Triton 1%). Reported values are means ± SEM of three independent experiments and p value was calculated using the Bonferroni multiple comparison test (** p < 0.01; *** p < 0.001) with untreated cells (CTRL).

Article Snippet: HSF cells were obtained from ScienCell research laboratory (ScienCell, 4700 Clinisciences) and were grown in Modified Eagle's Medium (MEM, PAN Biotech P0408500) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech, 3302 P290907), 2 mM of L-glutamine (Biochrom AG, K0282), 0.1 mg/mL penicillin–streptomycin (PAN Biotech, P0607100), 1 mM of sodium pyruvate (PAN Biotech, P0443100) and 0,5 μg/mL of amphotericin B (PAN Biotech, P0601001).

Techniques: Concentration Assay, Cell Culture, Infection, Cytotoxicity Assay, Lysis, Comparison

Journal: Scientific Reports

Article Title: Quercetin can reduce viral RNA level of O’nyong-nyong virus and resulting innate immune cytokine responses in cultured human synovial fibroblasts

doi: 10.1038/s41598-021-85840-z

Figure Lengend Snippet: Quercetin impairs HSF cells infection by ONNV. HSF cells were exposed to ONNV (MOI 1) co-treated or not with phenolics (10 µM) for 24 h. Cells were probed with primary antibody Alpha SC 58,088 anti-alphavirus before nuclei staining by DAPI (blue) and antibody binding by Alexa Fluor 488-conjugated donkey anti-mouse IgG antibody (green) ( A ). Untreated cells (CTRL) and infected cells (+ ONNV) ( B ) Alexa Fluor 488/DAPI overlay upon phenolic treatment. ( C ) Ratio Alexa Fluor 488 / DAPI fluorescence intensity with at least 10,000 DAPI stained cells. Reported values are means ± SEM of three independent experiments. p value was calculated using the Bonferroni multiple comparison test (*** p < 0.001) between infected cells (+ ONNV) and untreated cells (CTRL) or ( ## p < 0.01, ### p < 0.001) between infected cells (+ ONNV) and infected cells upon treatment with phenolics.

Article Snippet: HSF cells were obtained from ScienCell research laboratory (ScienCell, 4700 Clinisciences) and were grown in Modified Eagle's Medium (MEM, PAN Biotech P0408500) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech, 3302 P290907), 2 mM of L-glutamine (Biochrom AG, K0282), 0.1 mg/mL penicillin–streptomycin (PAN Biotech, P0607100), 1 mM of sodium pyruvate (PAN Biotech, P0443100) and 0,5 μg/mL of amphotericin B (PAN Biotech, P0601001).

Techniques: Infection, Staining, Binding Assay, Fluorescence, Comparison

Journal: Scientific Reports

Article Title: Quercetin can reduce viral RNA level of O’nyong-nyong virus and resulting innate immune cytokine responses in cultured human synovial fibroblasts

doi: 10.1038/s41598-021-85840-z

Figure Lengend Snippet: Quercetin impairs ONNV replication in HSF cells. HSF cells were infected by ONNV (MOI 1) co-treated or not with phenolic compounds (10 µM) for 24 h. Then, RNA was collected and E1, E2, nsP1 and nsP2 gene expression levels were determined by qRT-PCR. Reported values are means ± SEM of three independent experiments. p value was calculated using the Bonferroni multiple comparison test: *p < 0.05, **p < 0.01, ***p < 0.001 when compared to non-infected and non-treated cells (CTRL) and # p < 0.05, ## p < 0.01, ### p < 0.001 when compared to infected cells (+ ONNV).

Article Snippet: HSF cells were obtained from ScienCell research laboratory (ScienCell, 4700 Clinisciences) and were grown in Modified Eagle's Medium (MEM, PAN Biotech P0408500) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech, 3302 P290907), 2 mM of L-glutamine (Biochrom AG, K0282), 0.1 mg/mL penicillin–streptomycin (PAN Biotech, P0607100), 1 mM of sodium pyruvate (PAN Biotech, P0443100) and 0,5 μg/mL of amphotericin B (PAN Biotech, P0601001).

Techniques: Infection, Gene Expression, Quantitative RT-PCR, Comparison

Journal: Scientific Reports

Article Title: Quercetin can reduce viral RNA level of O’nyong-nyong virus and resulting innate immune cytokine responses in cultured human synovial fibroblasts

doi: 10.1038/s41598-021-85840-z

Figure Lengend Snippet: Quercetin impairs ONNV replication in supernatants in HSF cells. HSF cells were infected by ONNV (MOI 1) co-treated or not with phenolic compounds (10 µM) for 24 h. Then, RNA was collected in the supernatants and E1, E2, nsP1 and nsP2 gene expression levels were determined by qRT-PCR. Reported values are means ± SEM of three independent experiments. p value was calculated using the Bonferroni multiple comparison test: *p < 0.05, **p < 0.01, ***p < 0.001 when compared to non-infected and non-treated cells (CTRL) and # p < 0.05, ## p < 0.01 when compared to infected cells (+ ONNV).

Article Snippet: HSF cells were obtained from ScienCell research laboratory (ScienCell, 4700 Clinisciences) and were grown in Modified Eagle's Medium (MEM, PAN Biotech P0408500) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech, 3302 P290907), 2 mM of L-glutamine (Biochrom AG, K0282), 0.1 mg/mL penicillin–streptomycin (PAN Biotech, P0607100), 1 mM of sodium pyruvate (PAN Biotech, P0443100) and 0,5 μg/mL of amphotericin B (PAN Biotech, P0601001).

Techniques: Infection, Gene Expression, Quantitative RT-PCR, Comparison

Journal: Scientific Reports

Article Title: Quercetin can reduce viral RNA level of O’nyong-nyong virus and resulting innate immune cytokine responses in cultured human synovial fibroblasts

doi: 10.1038/s41598-021-85840-z

Figure Lengend Snippet: Quercetin reduces CCL-2 and IL-1β pro-inflammatory mediators production in ONNV infected HSF cells. HSF cells were infected by ONNV (MOI 1) and/or treated with phenolics (10 µM) for 24 h. Then, RNA was collected and CCL-2 and IL-1β gene expression levels were determined by qRT-PCR, ( A ) Pro-inflammatory mediators RNA levels. Then, culture media were collected CCL-2 and IL-1β levels evaluated by ELISA, ( B ) Pro-inflammatory mediators levels. Reported values are means ± SEM of three independent experiments. p value was calculated using the Bonferroni multiple comparison test: *p < 0.05, **p < 0.01, ***p < 0.001 when compared to non-infected and non-treated cells (CTRL) and # p < 0.05, ## p < 0.01, ### p < 0.001 when compared to infected cells (+ ONNV).

Article Snippet: HSF cells were obtained from ScienCell research laboratory (ScienCell, 4700 Clinisciences) and were grown in Modified Eagle's Medium (MEM, PAN Biotech P0408500) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech, 3302 P290907), 2 mM of L-glutamine (Biochrom AG, K0282), 0.1 mg/mL penicillin–streptomycin (PAN Biotech, P0607100), 1 mM of sodium pyruvate (PAN Biotech, P0443100) and 0,5 μg/mL of amphotericin B (PAN Biotech, P0601001).

Techniques: Infection, Gene Expression, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Comparison