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: Molecular Pharmaceutics

Article Title: Paired Agent Imaging to Evaluate the Improvement of Affinity-Matured Nimotuzumab K4 and K5 Variants as EGFR-Detecting Optical Imaging Agents

doi: 10.1021/acs.molpharmaceut.5c01261

Figure Lengend Snippet: Paired agent imaging (PAI). (A) Coinjection of nimotuzumab labeled with 800CW or 680RD NIR dyes in a 1:1 ratio into mice bearing A-431 xenografts. Mice are imaged in the 800 and 700 nm channels. Merge: Represents an overlay of the NIR fluorescence in the 800 and 700 nm channels. A ratio of the signal in the 800 nm:700 nm channel. S = stomach; T = tumor. (B) Coinjection of nimotuzumab-IRDye800CW and an untargeted control IgG-IRDye680RD administered at a 1:1 ratio into mice bearing A-431 xenografts. Mice are shown as a merge (overlay of the 800 and 700 nm channels) and a ratio of the 800 nm:700 nm fluorescence signal. (C) Coinjection of nimotuzumab labeled with 800CW or 680RD at a 2:1 ratio, respectively, into mice bearing A-431 xenografts. Mice are shown as a merge (overlay of the 800 and 700 nm channels) and a ratio of the 800 nm:700 nm fluorescence signal. (D) NIR fluorescent signal in the tumor is shown in MFI per area (mm 2 ) versus time, and (E) the ratio of 800 nm:700 nm NIR fluorescence signal versus time for nimotuzumab-IRDye800CW and nimotuzumab-IRDye680RD coinjected at a 1:1 and 2:1 ratio, respectively, and nimotuzumab-IRDye800CW and control IgG-IRDye680RD at a 1:1 ratio. The 95% confidence interval is shown in gray shading in D and E. C = control, Nimo = nimotuzumab.

Article Snippet: MDA-MB-468 and A-431 cells were purchased from ATCC and cultured in 90% Dulbecco’s Modified Eagle’s Medium (DMEM) or Roswell Park Memorial Institute (RPMI) medium, respectively, and 10% Fetal Bovine Serum (FBS) with 5% CO 2 in a humidified incubator.

Techniques: Imaging, Labeling, Fluorescence, Control

Journal: Molecular Pharmaceutics

Article Title: Paired Agent Imaging to Evaluate the Improvement of Affinity-Matured Nimotuzumab K4 and K5 Variants as EGFR-Detecting Optical Imaging Agents

doi: 10.1021/acs.molpharmaceut.5c01261

Figure Lengend Snippet: PAI of K4 or K5 relative to nimotuzumab in mice bearing MDA-MB-468 xenografts. (A) K4-IRDye800CW, K5-IRDye800CW, or nimotuzumab-IRDye800CW were coinjected with nimotuzumab-IRDye680RD into mice bearing MDA-MB-468 xenografts and imaged over time. The ratio of the fluorescent signal of K4-IRDye800CW:nimotuzumab-IRDye680RD (K4/Nimo), K5-IRDye800CW:nimotuzumab-IRDye680RD (K5/Nimo), and nimotuzumab-IRDye800CW:nimotuzumab-IRDye680RD (Nimo/Nimo) versus time is shown for each mouse. (B) The ratios of the MFI of K4/Nimo, K5/Nimo, and Nimo/Nimo in the A-431 xenograft tumor were normalized for labeling ratio and plotted versus time. The 95% confidence interval is shown in gray shading. (C) Correlation of K D app with the K4-IRDye800CW:nimotuzumab-IRDye680RD, K5-IRDye800CW:nimotuzumab-IRDye680RD, and nimotuzumab-IRDye800CW:nimotuzumab-IRDye680RD ratios of 800 nm:700 nm at 24, 48, 72, and 96 h postinjection. The error is the standard deviation.

Article Snippet: MDA-MB-468 and A-431 cells were purchased from ATCC and cultured in 90% Dulbecco’s Modified Eagle’s Medium (DMEM) or Roswell Park Memorial Institute (RPMI) medium, respectively, and 10% Fetal Bovine Serum (FBS) with 5% CO 2 in a humidified incubator.

Techniques: Labeling, Standard Deviation