magnetic separation kit  (Miltenyi Biotec)

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 GBC cell lines NOZ (CC-Y1668) and GBC-SD (CC-Y1162) were obtained from Shanghai EK-Bioscience Biotechnology Co., Ltd. Human monocytic cells THP-1 (TIB-202) and 293T cells (CRL-3216) were purchased from the ATCC.

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 GBC cell lines NOZ (CC-Y1668) and GBC-SD (CC-Y1162) were obtained from Shanghai EK-Bioscience Biotechnology Co., Ltd. Human monocytic cells THP-1 (TIB-202) and 293T cells (CRL-3216) were purchased from the ATCC.

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

Journal: Cell Insight

Article Title: TNF inhibits SARS-CoV-2 induced cell-cell fusion through activating the SDC4-RhoA signaling to promote actin bundles formation

doi: 10.1016/j.cellin.2026.100310

Figure Lengend Snippet: Host factors secreted by innate immune cells early after activation inhibit SARS-CoV-2 spike protein induced cell-cell fusion. (A) Luciferase assay showing the effect of supernatant from THP-1 cell cultures pretreated with the NLRP3 inhibitor MCC950 (10 μM) and then stimulated with the TLR1/2 ligand Pam3CSK4 (1 μg/mL) for 3 h on spike protein-induced HEK293T cell-cell fusion. Serum-free RPMI 1640 served as the medium control. Data points represent mean ± SEM from six independent experiments; P values are indicated. (B) Immunoblot analysis of S2′ cleavage in fused cells treated with supernatant from MCC950-pretreated THP-1 cultures stimulated with Pam3CSK4 for 3 h. Blots are representative of three independent experiments. (C) Luciferase assay showing the effect of supernatant from NLRP3-knockout THP-1 cell cultures stimulated with Pam3CSK4 for 3 h on spike-induced HEK293T cell-cell fusion. Serum-free RPMI 1640 served as the control. Data points represent mean ± SEM from six independent experiments; P values are indicated. (D) Immunoblot analysis of S2′ cleavage in fused cells treated with supernatant from NLRP3-knockout THP-1 cell cultures stimulated with Pam3CSK4 for 3 h. Blots are representative of three independent experiments. (E) Visualization of syncytium formation by ZsGreen fluorescence after treatment with supernatant from MCC950-pretreated THP-1 cultures stimulated with Pam3CSK4 for 3 h. Images are representative of three independent experiments; white arrows indicate syncytia. Scale bar, 50 μm. (F) Visualization of syncytium formation by ZsGreen fluorescence after treatment with supernatant from NLRP3-knockout THP-1 cell cultures stimulated with Pam3CSK4 for 3 h. Images are representative of three independent experiments; white arrows indicate syncytia. Scale bar, 50 μm. (G) Quantification of relative syncytium area (%) based on ZsGreen fluorescence images in (E). Data are presented as mean ± SEM from three independent experiments. P values are indicated above the bars. (H) Quantification of relative syncytium area (%) based on ZsGreen fluorescence images in (F). Data are presented as mean ± SEM from three independent experiments. P values are indicated.

Article Snippet: The human monocytic THP-1 cell line (TIB-202; ATCC) was authenticated via short tandem repeat (STR) analysis by Suzhou Genetic Testing Biotech Co., Ltd, following the ANSI/ATCC ASN-0002-2012 standard ( ; ).

Techniques: Activation Assay, Luciferase, Control, Western Blot, Knock-Out, Fluorescence

Journal: Cell Insight

Article Title: TNF inhibits SARS-CoV-2 induced cell-cell fusion through activating the SDC4-RhoA signaling to promote actin bundles formation

doi: 10.1016/j.cellin.2026.100310

Figure Lengend Snippet: TNF produced by innate immune cells early after activation inhibit SARS-CoV-2 spike induced cell-cell fusion. (A) Luciferase assay showing the effect of recombinant IL-6 (10 ng/mL), IL-8 (10 ng/mL), or TNF (10 ng/mL) on spike-induced cell-cell fusion. PBS was used as the vehicle control. Data points represent mean ± SEM from four independent experiments; P values are indicated. (B) Luciferase assay showing the effect of supernatant from THP-1 cultures stimulated with Pam3CSK4 for the indicated durations on cell-cell fusion in HEK293T cells pretreated with the IL-1 receptor antagonist (IL-1RA) (4 μg/mL). Data represent mean ± SEM from four independent experiments; P values are shown. (C) Immunoblot analysis of S2′ cleavage in IL-1RA-pretreated fused cells exposed to supernatant from THP-1 cultures stimulated with Pam3CSK4 for the indicated durations. Blots are representative of three independent experiments. (D) ELISA quantification of IL-1β and TNF levels in supernatant from THP-1 cell cultures after stimulation with the indicated TLR ligand at the specified time points. Data represent mean ± SEM from three independent experiments. (E) Luciferase assay showing the effect of supernatant from THP-1 cultures stimulated with Pam3CSK4 for the indicated durations on spike-induced fusion in HEK293T-sgcontrol and HEK293T-sgTNFR1 cells. Data points represent mean ± SEM from six independent experiments; P values are indicated. (F) Immunoblot analysis of S2′ cleavage in HEK293T-sgcontrol and HEK293T-sgTNFR1 fused cells treated with supernatant from THP-1 cell cultures stimulated with Pam3CSK4 for the indicated durations. Blots are representative of three independent experiments. (G) Luciferase assay showing the effect of supernatant from Pam3CSK4-stimulated THP-1 cell cultures on cell-cell fusion in IL-1RA-pretreated HEK293T-sgcontrol and HEK293T-sgTNFR1 cells. Data represent mean ± SEM from four independent experiments; P values are indicated. (H) Immunoblot analysis of S2′ cleavage in IL-1RA-pretreated HEK293T-sgcontrol and HEK293T-sgTNFR1 fused cells exposed to supernatant from Pam3CSK4-stimulated THP-1 cell cultures. Blots are representative of three independent experiments. (I–J) ELISA quantification of IL-1β and TNF levels in supernatant from THP-1 cell cultures under (I) MCC950 pharmacological inhibition of NLRP3, (J) NLRP3 knockout, after stimulation with the indicated TLR ligand at the specified time points. Data represent mean ± SEM from three independent experiments.

Article Snippet: The human monocytic THP-1 cell line (TIB-202; ATCC) was authenticated via short tandem repeat (STR) analysis by Suzhou Genetic Testing Biotech Co., Ltd, following the ANSI/ATCC ASN-0002-2012 standard ( ; ).

Techniques: Produced, Activation Assay, Luciferase, Recombinant, Control, Western Blot, Enzyme-linked Immunosorbent Assay, Inhibition, Knock-Out