Journal: Synthetic and Systems Biotechnology

Article Title: Leveraging ANXA1 to enhance recombinant protein yields in CHO cells: A UPR-Mediated bioprocessing approach

doi: 10.1016/j.synbio.2025.12.001

Figure Lengend Snippet: Schematic overview of the experimental workflow for generating ANXA1-knockdown CHO cell lines and AC2-26 inhibitor treatment. (A) Generation and validation of ANXA1-knockdown cell lines for rADM antibody production. (B) AC2-26 inhibitor treatment in low-producer CHO cells (ADM-14) and subsequent rADM expression analysis.

Article Snippet: On day 3 of suspension culture, the small molecule inhibitor AC2-26 (Topscience Co., Ltd., China) was added [ ], using DMSO (Solarbio Life Sciences, China) as the solvent control.

Techniques: Knockdown, Biomarker Discovery, Expressing

Journal: Synthetic and Systems Biotechnology

Article Title: Leveraging ANXA1 to enhance recombinant protein yields in CHO cells: A UPR-Mediated bioprocessing approach

doi: 10.1016/j.synbio.2025.12.001

Figure Lengend Snippet: AC2-26 effect on ADM-14 CHO cells. (A) Cell density/viability under AC2-26 treatment (n = 3). (B) rADM expression by Western blot with quantification (n = 3). (C) ANXA1 mRNA/protein comparison (n = 3). Quantification was performed using ImageJ (for Western blot densitometry) and GraphPad Prism 10 (for statistical analysis). (∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001). n: represents independent biological replicates.

Article Snippet: On day 3 of suspension culture, the small molecule inhibitor AC2-26 (Topscience Co., Ltd., China) was added [ ], using DMSO (Solarbio Life Sciences, China) as the solvent control.

Techniques: Expressing, Western Blot, Comparison

Journal: Cellular Oncology

Article Title: Upregulation of limb-bud and heart (LBH) drives liver cancer progression by interacting with the oncoprotein Mortalin

doi: 10.1007/s13402-026-01228-z

Figure Lengend Snippet: LBH is transcriptionally upregulated by TGF-β in liver cancer cells. (A) Heatmap displaying the common 474 DEGs whose expressions were altered upon treatment with 100 pM of TGF-β1 for 4 h in three different HCC cell lines, i.e. MHCC97H, MHCC97L and HCCLM3. (B) Heatmap classification displaying some typical upregulated genes in response to TGF-β1 administration in HCC cell lines. (C-H) After being treated with 100 pM of TGF-β1 (C-E) or 10 µM of SB431542 (F-H) for the indicated periods, HCCLM3, HLE and SMMC-7721 cells were harvested for RNA purification and gene expression analyses through qPCR. (I-J) HCCLM3, HLE cells transfected with a control siRNA (NS) or those targeting Smad2/3 were treated with 100 pM of TGF-β1 for 6 h. Then the expression levels of Smad2, Smad3 and LBH were analyzed by qPCR. (K) Gene track view of Smad2/3, H3K4me3, and H3K27ac at the LBH promoter from the Cistrome Data database (top), and schematic diagram of ChIP primers designed within and near the LBH promoter region (bottom). HCCLM3 cells were treated with 2.5 ng/ml TGF-β for 24 h. ChIP assays were then performed to examine the binding of Smad2/3 to the LBH promoter region, and the enriched DNA fragments were quantitatively analyzed by qPCR. qPCR experiments described above were analyzed by one-way ANOVA followed by Tukey’s or Dunnett’s post hoc test. Statistical significance: * p < 0.05, ** p < 0.01 and *** p < 0.001; ns, no significance. Data were shown as mean ± SD

Article Snippet: Recombinant human TGF-β1 protein was obtained from R&D Systems Inc., and the small molecule inhibitor SB431542 was purchased from MedChemExpress (MCE, USA).

Techniques: Purification, Gene Expression, Transfection, Control, Expressing, Binding Assay

Journal: Cancers

Article Title: Paquinimod Targeting of the S100A8/A9 Axis Suppresses Liver Metastasis in Aged Mice

doi: 10.3390/cancers18101635

Figure Lengend Snippet: Association of S100A9 inhibition with lower tumor burden and altered immune cell composition in aged metastatic livers. ( A ) Experimental schema of tumor cell inoculation and every other day paquinimod or vehicle administration in young and aged mice. ( B ) Representative IVIS bioluminescence images of liver metastases at days 14 and 21 after tumor inoculation (left). Comparisons of tumor burden (radiance) between day 14 and day 21 within each treatment group are shown (upper right); paired t -tests were performed (Young: n = 3 per group; Aged: n = 7 per group). The change in tumor burden (Δlog10 photons/sec/cm 2 /sr) from day 14 to day 21 is summarized (lower right) for young and aged mice. For the Δlog10 bar graphs, vehicle- and paquinimod-treated mice were compared using unpaired Student’s t -tests (n = 3 young and n = 7 aged). ( C ) Representative macroscopic appearance of livers (left) and quantification of the number of metastatic nodules and the tumor area ratio (right) at day 21 (n = 7 per group). ( D – F ) Representative flow cytometry plots and frequencies of tumor-infiltrating immune cells at day 21 after tumor inoculation, including Ly6G + cells, Ly6C + monocytes, PMN-MDSCs (Ly6G + CD244 + ), CD4 + and CD8 + T cells, and NK cells. Absolute numbers are shown alongside frequencies (n = 3 per group). ( G ) Correlation between day 21 tumor burden and frequencies of the indicated immune cell populations in aged mice, assessed by simple linear regression; R 2 and p values are shown. (n = 14 aged mice). Data are presented as mean ± SEM. Statistical analyses were performed using Student’s t -test, paired t -test, or the Mann–Whitney U test, as appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: S100A9 inhibition studies were performed using the small-molecule inhibitor Paquinimod (HY-100442; MedChemExpress, Monmouth Junction, NJ, USA), also known as ABR215757 [ , ].

Techniques: Inhibition, Flow Cytometry, MANN-WHITNEY

Journal: bioRxiv

Article Title: Sortilin deficiency alters baseline retinal homeostasis and injury-induced signaling without affecting optic nerve crush-induced neurodegeneration

doi: 10.64898/2026.05.08.723723

Figure Lengend Snippet: A. Experimental plan: Following baseline optical coherence tomography (OCT) scans, C57BL6/JRj mice were subjected to optic nerve crush (ONC) in one eye and sham procedure in the other eye. Immediately after, mice were randomized to receive intravitreal (IVT) injection of either the anti-sortilin polyclonal antibody (pAb) AF2934 or IgG control (2 µL of 1 µg/µL) in the crush eye while the other served as untreated sham control. Mice were sacrificed following OCT at 14 days post crush (dpc). B. Bar plot (mean ± sd) showing quantification of the change from baseline of the combined thickness of the nerve fiber, ganglion cell and inner nuclear layer, denoted the NGI thickness. Statistical comparisons were performed using one-way ANOVA followed by Tukey’s test (n > 5). C. Bar plot (mean ± sd) showing quantification of the density of RNA-binding protein with multiple splicing (RbPMS) positive retinal ganglion cells (RGCs) in retinal flat mounts. Statistical comparisons were performed by one-way ANOVA followed by Tukey’s test (n > 5). Representative OCT scans (D) and areas from anti-RbPMS (red) immunostained retinal flat mounts (E) are shown. Scale bars 50 µm. F. Experimental plan: As in (A), C57BL6/JRj mice were randomized to receive IVT injection of the small-molecule inhibitor AF38469 (2 µL of 1 µg/µL) or vehicle (PBS-1% DMSO) in the crush eye. G. Bar plot (mean ± sd) showing quantification of the change from baseline of NGI thickness. Statistical comparisons were performed by one-way ANOVA followed by Tukey’s test (n > 5). H. Bar plot (mean ± sd) showing quantification of RGC density in retinal flat mounts. Statistical comparisons were performed by one-way ANOVA followed by Tukey’s test (n > 5). Representative OCT scans (I) and areas from anti-RbPMS (red) immunostained retinal flat mounts (J) are shown. Scale bars 50 µm. Significance levels: * [0.01, 0.05]; ** [0.001,0.01]; *** [0.0001, 0.001]; **** [0, 0.0001].

Article Snippet: In the second experiment, the small-molecule inhibitor AF38469 (MedChemExpress LLC, Monmouth Junction, NJ, USA) was compared with the buffer solution.

Techniques: Tomography, Injection, Control, RNA Binding Assay