Journal: The Journal of Biological Chemistry

Article Title: Hyaluronic acid regulates cellular UDP-GlcNAc levels through CD44 to affect glycosylation and cell biological functions

doi: 10.1016/j.jbc.2025.111111

Figure Lengend Snippet: Establishment of HAS2 KO cells and confirmation of HA synthesis. A , comparison of HAS2 expression levels among different cell lines: HeLa (cervical adenocarcinoma cell line), A431 (epidermoid carcinoma cell line), PANC-1 (pancreatic ductal adenocarcinoma cell line), MIA PaCa-2 (pancreatic carcinoma cell line), and HepG2 (hepatocellular carcinoma cell line). The HAS2 mRNA level in HeLa was normalized to 1.0. All values are presented as mean ± SD from three independent experiments, based on one-way ANOVA with Tukey’s post hoc analysis. ∗∗∗ p < 0.001. B , comparison of HAS1 , HAS2 , and HAS3 expression levels in HeLa cells. The HAS1 mRNA level in HeLa was normalized to 1.0. All values are presented as mean ± SD from three independent experiments, based on one-way ANOVA with Tukey’s post hoc analysis. ∗∗∗ p < 0.001. C , Sanger sequencing was used to analyze exon 2 of the target sequence in HeLa WT and HAS2 KO cells with the canonical SpCas9 PAM sequence (TGG) highlighted in bold. D , WT and HAS2 KO cells were cultured in serum-free medium, and the conditioned medium was collected after 24 h to measure HA concentration. Statistical significance was determined from three independent experiments using an unpaired Student’s t test, with p values indicated as ∗∗∗ p < 0.001.

Article Snippet: A431 cells were obtained from ATCC.

Techniques: Comparison, Expressing, Sequencing, Cell Culture, Concentration Assay

Journal: bioRxiv

Article Title: IL-1α drives a tumor-stroma-neutrophil axis through inflammatory fibroblast activation in head and neck cancer

doi: 10.64898/2026.01.20.700440

Figure Lengend Snippet: (A) MSCs were stimulated for 24 h with FaDu tumor-conditioned SNs. Tumor-derived cytokines (columns) were quantified by Luminex, and MSC-derived CXCL8 and G-CSF (rows) by ELISA. Data are displayed as a correlation matrix. (B) MSCs were treated with FaDu tumor SNs for 24 h. Tumor-derived factors were analyzed by Luminex; MSC-derived CXCL8 and G-CSF were quantified by ELISA. Tumor-derived IL-1α correlates with MSC-derived CXCL8 and G-CSF. (C) MSCs were treated with recombinant IL-1α for 24 h. Release of CXCL8 and G-CSF was analyzed by ELISA. (D) IL-1α was measured in control (non-sense, NS) and IL-1α overexpressing FaDu cells (IL-1α-OE) using Luminex. (E) MSCs were treated with SNs from non-sense and IL-1α-OE cells. MSC-derived CXCL8 and G-CSF were quantified by ELISA. (F) IL-1α release was determined in the SN of viable and necrotic FaDu and A431 cells. MSCs were treated with the SN of viable and necrotic FaDu (G) or A431 (H) cells for 24 h. MSC-derived CXCL8 and G-CSF were quantified by ELISA. Statistical significance was assessed after log-transformation using an ordinary one-way ANOVA with Tukey’s multiple comparisons test (C), while paired t -tests were applied for panels D-H. Data are shown as mean ± SD. In panel C, significance levels are indicated as # or * (p ≤ 0.05), ## or ** (p ≤ 0.01), ### or *** (p ≤ 0.001), and #### or **** (p ≤ 0.0001); all other p values are shown numerically. Symbols (BioRender) are included to show the origin of the analyzed SNs.

Article Snippet: Human carcinoma cell lines: The epidermoid carcinoma cell line A431 (ATCC CRL-1555) was cultured in DMEM with 4.5 g/L glucose (Pan-Biotech, Aidenbach, Germany), supplemented with 10% heat inactivated FCS (BioSell, Feucht, Germany) and 100 units/ml penicillin and 100 μg/mL streptomycin (Thermo Fisher Scientific, Waltham, USA).

Techniques: Derivative Assay, Luminex, Enzyme-linked Immunosorbent Assay, Recombinant, Control, Transformation Assay

Journal: bioRxiv

Article Title: IL-1α drives a tumor-stroma-neutrophil axis through inflammatory fibroblast activation in head and neck cancer

doi: 10.64898/2026.01.20.700440

Figure Lengend Snippet: (A,B) Tumor-derived cytokines were quantified by Luminex analysis. (A) Data are displayed as Pearsońs correlation matrix. Significant correlations among tumor-derived factors are indicated by black asterisks (B). Quantification of tumor-derived mediators known to be involved in tumor-stroma communication. (C) MSCs were treated with FaDu-CXCL8KO tumor conditioned medium in the presence of 10 µg/mL IL-1α neutralizing antibody or isotype control. Released CXCL8 and G-CSF were quantified by ELISA. (D) Quantification of tumor-derived mediators of tumor-stroma communication in non-sense and IL-1α overexpressing (IL-1α-OE) cells was performed by Luminex. (E) Quantification of tumor-derived factors implicated in neutrophil recruitment and survival in SN from non-sense and IL-1α-OE cells (Luminex and CXCL8-ELISA). (F) LDH assay quantifying cytotoxicity in SN of viable and necrotic FaDu and A431 tumor cells. Statistical analysis was performed using paired t -tests (C,F) and unpaired t -tests (D). Data are presented as mean ± SEM (C) and mean ± SD (D,E), p values are indicated.

Article Snippet: Human carcinoma cell lines: The epidermoid carcinoma cell line A431 (ATCC CRL-1555) was cultured in DMEM with 4.5 g/L glucose (Pan-Biotech, Aidenbach, Germany), supplemented with 10% heat inactivated FCS (BioSell, Feucht, Germany) and 100 units/ml penicillin and 100 μg/mL streptomycin (Thermo Fisher Scientific, Waltham, USA).

Techniques: Derivative Assay, Luminex, Control, Enzyme-linked Immunosorbent Assay, Lactate Dehydrogenase Assay

Journal: Scientific Reports

Article Title: PISTOL MRI allows noninvasive assessment of changes in tumor oxygenation post hypoxia targeted therapy

doi: 10.1038/s41598-025-32618-2

Figure Lengend Snippet: Comparing baseline oxygenation (pO 2 ), edema fraction and tumor response to tirapazamine (TPZ) treatment in A431 and H1975 tumors. Baseline (pre-treatment, day 0) ( A ) oxygenation of the entire tumor and its center and peripheral regions and ( B ) edema fraction compared between A431 and H1975 tumors. Tumor volume of treated and untreated ( C ) A431 and (D) H1975 tumors over 15 days post treatment. Note: * represents the statistical difference between A431 and H1975 tumors. (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Human NCI-H1975 non-small cell lung cancer and A431 epidermoid carcinomas cells (ATCC Inc.) were cultured in Roswell Park Memorial Institute (RPMI 1640) and Dulbecco’s Modified Eagle’s Medium (DMEM) respectively, supplemented with 1% penicillin/streptomycin and 10% fetal bovine serum (Life Technologies, Carlsbad, CA, USA), in a 5% CO 2 -containing humidified atmosphere at 37 ° C. Immunocompromised male nu\nu mice (5–11/per cohort, Charles River Laboratories, USA) were implanted subcutaneously with 2 × 10 6 cells/40 μl of the respective cell line (A431 or NCI-H1975) in the right thigh.

Techniques:

Journal: Scientific Reports

Article Title: PISTOL MRI allows noninvasive assessment of changes in tumor oxygenation post hypoxia targeted therapy

doi: 10.1038/s41598-025-32618-2

Figure Lengend Snippet: Overlay of the reference T 2 -weighted tumor image and the respective pO 2 map from day 0 – 15 for a representative tumor from the control ( a - d ) and TPZ ( e - h )) treated A431 cohorts, respectively. The region of interest (shown as red boundary) represents the segmentation that was conducted to delineate the tumor from surrounding tissue and to segment the tumor into center and periphery. Numbers represent mean pO 2 values ± standard deviation in torr.

Article Snippet: Human NCI-H1975 non-small cell lung cancer and A431 epidermoid carcinomas cells (ATCC Inc.) were cultured in Roswell Park Memorial Institute (RPMI 1640) and Dulbecco’s Modified Eagle’s Medium (DMEM) respectively, supplemented with 1% penicillin/streptomycin and 10% fetal bovine serum (Life Technologies, Carlsbad, CA, USA), in a 5% CO 2 -containing humidified atmosphere at 37 ° C. Immunocompromised male nu\nu mice (5–11/per cohort, Charles River Laboratories, USA) were implanted subcutaneously with 2 × 10 6 cells/40 μl of the respective cell line (A431 or NCI-H1975) in the right thigh.

Techniques: Control, Standard Deviation

Journal: Scientific Reports

Article Title: PISTOL MRI allows noninvasive assessment of changes in tumor oxygenation post hypoxia targeted therapy

doi: 10.1038/s41598-025-32618-2

Figure Lengend Snippet: Changes in tumor pO 2 and statistical comparison of all A431 cohorts ( A - F ). Note: no significant difference was observed compared to baseline values for any cohort or time point).

Article Snippet: Human NCI-H1975 non-small cell lung cancer and A431 epidermoid carcinomas cells (ATCC Inc.) were cultured in Roswell Park Memorial Institute (RPMI 1640) and Dulbecco’s Modified Eagle’s Medium (DMEM) respectively, supplemented with 1% penicillin/streptomycin and 10% fetal bovine serum (Life Technologies, Carlsbad, CA, USA), in a 5% CO 2 -containing humidified atmosphere at 37 ° C. Immunocompromised male nu\nu mice (5–11/per cohort, Charles River Laboratories, USA) were implanted subcutaneously with 2 × 10 6 cells/40 μl of the respective cell line (A431 or NCI-H1975) in the right thigh.

Techniques: Comparison

Journal: Scientific Reports

Article Title: PISTOL MRI allows noninvasive assessment of changes in tumor oxygenation post hypoxia targeted therapy

doi: 10.1038/s41598-025-32618-2

Figure Lengend Snippet: Edema fraction of A431 and H1975 treatment groups. Treatment-induced changes in edema fraction in ( A ) A431 and ( B ) H1975 tumors. Post-treatment differences in edema fractions between ( C ) A431 and ( D ) H1975 tumors. (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Human NCI-H1975 non-small cell lung cancer and A431 epidermoid carcinomas cells (ATCC Inc.) were cultured in Roswell Park Memorial Institute (RPMI 1640) and Dulbecco’s Modified Eagle’s Medium (DMEM) respectively, supplemented with 1% penicillin/streptomycin and 10% fetal bovine serum (Life Technologies, Carlsbad, CA, USA), in a 5% CO 2 -containing humidified atmosphere at 37 ° C. Immunocompromised male nu\nu mice (5–11/per cohort, Charles River Laboratories, USA) were implanted subcutaneously with 2 × 10 6 cells/40 μl of the respective cell line (A431 or NCI-H1975) in the right thigh.

Techniques:

Journal: Scientific Reports

Article Title: PISTOL MRI allows noninvasive assessment of changes in tumor oxygenation post hypoxia targeted therapy

doi: 10.1038/s41598-025-32618-2

Figure Lengend Snippet: Representative PIM (green) stained tumor sections from ( A ) A431 and ( B ) H1975 tumors (counterstained with DAPI). ( C ) The mean hypoxia fractions of the A431 and H1975 cohorts. TPZ treatment of H1975 tumors resulted in a higher extent of hypoxia compared to untreated control tumors. ( D & E ) No difference was observed between the center and periphery hypoxia fractions for each treatment group. (* p < 0.05).

Article Snippet: Human NCI-H1975 non-small cell lung cancer and A431 epidermoid carcinomas cells (ATCC Inc.) were cultured in Roswell Park Memorial Institute (RPMI 1640) and Dulbecco’s Modified Eagle’s Medium (DMEM) respectively, supplemented with 1% penicillin/streptomycin and 10% fetal bovine serum (Life Technologies, Carlsbad, CA, USA), in a 5% CO 2 -containing humidified atmosphere at 37 ° C. Immunocompromised male nu\nu mice (5–11/per cohort, Charles River Laboratories, USA) were implanted subcutaneously with 2 × 10 6 cells/40 μl of the respective cell line (A431 or NCI-H1975) in the right thigh.

Techniques: Staining, Control