Journal: Materials Today Bio

Article Title: Targeting FN1 to overcome gemcitabine resistance in gallbladder cancer: Mechanistic insights and an iRGD-modified PEG-PLGA nanoparticle delivery strategy

doi: 10.1016/j.mtbio.2026.102877

Figure Lengend Snippet: Effect of FN1 knockdown on the immunosuppressive microenvironment in GBC-SD/GEM cells. Note: (A) Schematic of immunosuppressive cells and cytokines detection in tumor tissues; (B) FCM analysis of Tregs infiltration levels in GBC-SD/GEM cell xenograft tissues in various mouse groups; (C) FCM analysis of M2 and M1 macrophage infiltration levels in GBC-SD/GEM cell xenograft tissues from different mouse groups; (D-E) RT-qPCR analysis of immunosuppressive factors expression in GBC-SD/GEM cell xenograft tissues from various mouse groups; (F) Schematic of in vitro co-culture of GEM-resistant GBC cells with THP-1 and CD4 + T cells; (G) FCM analysis of Tregs levels in CD4 + T cells after co-culture with GBC-SD/GEM cells; (H) FCM analysis of M2 and M1 macrophage levels in THP-1 cells post co-culture with GBC-SD/GEM cells; (I) RT-qPCR analysis of IL-10 or CSF-1 expression in CD4 + T or THP-1 cells after co-culture with GBC-SD/GEM cells. In B-E, ∗ indicates p < 0.05 compared to the sh-NC + GEM group, each group consisting of 6 mice; in G-I, ∗ indicates p < 0.05 compared to the sh-NC group, # indicates p < 0.05 compared to the oe-NC group, experiments repeated three times.

Article Snippet: Human CD4 + T cells were isolated and enriched from mature lymphocytes (hPBMCs, PCS-800-011, ATCC, USA) using a Human CD4 + T Cell Enrichment Kit (8804-6814-74, Thermo Fisher Scientific, USA). hPBMCs were incubated with the enrichment reagents for 30 min to capture CD4 + T cells.

Techniques: Knockdown, Quantitative RT-PCR, Expressing, In Vitro, Co-Culture Assay

Journal: Materials Today Bio

Article Title: Targeting FN1 to overcome gemcitabine resistance in gallbladder cancer: Mechanistic insights and an iRGD-modified PEG-PLGA nanoparticle delivery strategy

doi: 10.1016/j.mtbio.2026.102877

Figure Lengend Snippet: Impact of NPs delivering si-FN1 on drug resistance and immune cell infiltration in GBC-SD/GEM cells. Note: (A) Schematic of the experimental setup for studying the impact of NPs delivering si-FN1 on GBC GEM resistance; (B-C) RT-qPCR (B) and Western Blot (C) analysis of FN1 and PI3K pathway protein expression in GBC-SD/GEM cells treated with NPs (si-FN1) and iRGD-NPs (si-FN1); (D) CCK-8 assay assessing the viability changes in GBC-SD/GEM cells after treatment with NPs (si-FN1) and iRGD-NPs (si-FN1); (E) Clonogenic assay evaluating colony formation in various groups of GBC-SD/GEM and NOZ/GEM cells; (F) FCM analysis of apoptosis in GBC-SD/GEM cells across different groups; (G) FCM analysis of Tregs levels in CD4 + T cells after co-culture with GBC-SD/GEM cells; (H) FCM analysis of M2 and M1 macrophage levels in THP-1 cells after co-culture with GBC-SD/GEM cells; (I) RT-qPCR analysis of IL-10 or CSF-1 expression in CD4 + T or THP-1 cells co-cultured with GBC-SD/GEM cells. ∗ indicates p < 0.05 compared to the NPs (si-NC) group, # indicates p < 0.05 compared to the NPs (si-FN1) group, experiments repeated three times.

Article Snippet: Human CD4 + T cells were isolated and enriched from mature lymphocytes (hPBMCs, PCS-800-011, ATCC, USA) using a Human CD4 + T Cell Enrichment Kit (8804-6814-74, Thermo Fisher Scientific, USA). hPBMCs were incubated with the enrichment reagents for 30 min to capture CD4 + T cells.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, CCK-8 Assay, Clonogenic Assay, Co-Culture Assay, Cell Culture

Journal: Blood Neoplasia

Article Title: Olfactory receptors as tumor suppressors in cutaneous T-cell lymphoma via p38γ pathway modulation

doi: 10.1016/j.bneo.2025.100101

Figure Lengend Snippet: p38γ inhibitors increase olfactory transduction pathway activity. (A) WB analysis of Hut78 cells treated with 0.5 to 10 μM of CSH18. (B) WB analysis of Hut78 cells treated with 0.5 to 10 μM of CSH71. Both CSH18 and CSH71 increased OR4N5 protein levels. Other olfactory factors OR10K2, OR2H2, and OR4K17 were also assessed, with dimerization of OR4N5 observed based on molecular weight (blue arrows indicate 50 kDa). (C) WB assays for lentiviral expression of sh-p38γ vs sh-Ctrl. OR4N5 bands at ∼30 and 60 kDa correspond to the monomer and dimer, respectively (sh-Ctrl lane). (D) OR4N5 3D structure prediction by AlphaFold (AF-Q8IXE1-F1, left). Right: localization of OR4N5 in the plasma membrane (green, confidence score = 5). Slight presence in the cytoskeleton (confidence score = 1) according to Genecard.com . (E) In-depth analysis of the OR4N5 amino acid sequence identified 7 transmembrane domains (underlined) and 2 cysteines that may contribute to OR4N5 dimerization. CDR, Complementarity Determining Region; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; hr, hour; sh-Ctrl, shRNAs targeting control; sh-p38γ, shRNAs targeting p38γ; TM, Transmembrane.

Article Snippet: Previously, we showed that CSH71 (250 nM) strongly upregulates the olfactory transduction pathway in Hut78 cells (CD4 + , SS; American Type Culture Collection) via RNA microarray.

Techniques: Transduction, Activity Assay, Molecular Weight, Expressing, Clinical Proteomics, Membrane, Sequencing, Control

Journal: Blood Neoplasia

Article Title: Olfactory receptors as tumor suppressors in cutaneous T-cell lymphoma via p38γ pathway modulation

doi: 10.1016/j.bneo.2025.100101

Figure Lengend Snippet: Gene silencing of either OR4N5 or OR10K2 promotes cell proliferation in Hut78 cells. (A) Cell viability assays were conducted on OR4N5- and OR10K2-silenced cells (sh- OR4N5 and sh- OR10K2 ) over a 10-day period after transduction to assess cell proliferation. Cell counts were measured on days 1 through 10 using the trypan blue exclusion method to distinguish viable cells. A scramble sequence shRNA served as the control in these assays. (B) Confocal immunofluorescence analysis confirmed the successful silencing of OR4N5 and OR10K2 proteins in Hut78 cells (left). The loss of OR expression in Hut78 cells correlates with increased cell proliferation 7 to 9 days after transduction. Statistical data are presented as mean (standard deviation). Cells were counted on days 1, 4, 7, and 10 after selection using an automated cell counter, with trypan blue exclusion to differentiate viable cells. sh-Ctrl, short hairpin control.

Article Snippet: Previously, we showed that CSH71 (250 nM) strongly upregulates the olfactory transduction pathway in Hut78 cells (CD4 + , SS; American Type Culture Collection) via RNA microarray.

Techniques: Transduction, Sequencing, shRNA, Control, Immunofluorescence, Expressing, Standard Deviation, Selection

Journal: Blood Neoplasia

Article Title: Olfactory receptors as tumor suppressors in cutaneous T-cell lymphoma via p38γ pathway modulation

doi: 10.1016/j.bneo.2025.100101

Figure Lengend Snippet: Colocalization of OR4N5 and CD3E, and colocalization of p38γ with CD3Z in TCR complexes using confocal immunofluorescence microscopy. (A) Colocalization of OR4N5 and CD3E in PBMCs from healthy donors. (B) Colocalization of OR4N5 and CD3E in CTCL Hut78 cells, both treated with CSH71 (250 nM) and untreated controls. In both panels A and B, the interaction between CD3E and OR4N5 was assessed, with colocalization indicated by the yellow signal in merged 2-color images.

Article Snippet: Previously, we showed that CSH71 (250 nM) strongly upregulates the olfactory transduction pathway in Hut78 cells (CD4 + , SS; American Type Culture Collection) via RNA microarray.

Techniques: Immunofluorescence, Microscopy