Journal: bioRxiv

Article Title: SIRPA suppresses integrin-dependent virus endocytosis

doi: 10.64898/2026.04.17.719277

Figure Lengend Snippet: A) shSIRPA or shCTRL cells were treated with BTT-3033 and infected with the indicated viruses. RNA was isolated at 24 hpi and viral RNA was measured by RT-qPCR. Shown is the average ± SD of 3–4 independent experiments. One-way ANOVA was used to determine significance. *, P ≤ 0.05; ***, P ≤ 0.001; ****, P ≤ 0.0001. B) VIAs were carried out with U2OS cells treated with BTT-3033. Internalized viral RNA was analyzed by RT-qPCR. Shown is the average ± SD of 3 independent experiments. Student’s T test was used to determine significance. **, P ≤ 0.01; ****, P ≤ 0.0001. C) shSIRPA or shCTRL cells were treated with BTT-3033 and infected with JUNV C1. Viral titers were analyzed at 24 hpi. Shown is the average ± SD of 3 independent experiments. One-way ANOVA was used to determine significance. ns, not significant, **, P ≤ 0.01; ****, P ≤ 0.0001. D) U2OS cells were treated with BTT-3033 at indicated concentrations and infected with MACV or VSV pseudoviruses. Luciferase assays were performed at 48 hpi. Shown is the average ± SD of 3 independent experiments. One-way ANOVA was used to determine significance. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. E) Viral titers in serum and spleen of BTT-3033– and favipravir-treated mice. Horizontal lines represent the mean for each group. The dotted horizontal lines represent the assay’s lower limit of detection. * , P ≤ 0.05; ** , P ≤ 0.01; *** , P ≤ 0.001 compared between groups. One animal in the DMSO placebo group was excluded from the analysis due to complications unrelated to the viral infection.

Article Snippet: BTT-3033 and FAK inhibitor 14 (FIB-14) were purchased from TOCRIS, LHF-535 and favipiravir (6-fluoro-3-hydroxy-2-pyrazinecarboxamide) from TargetMol Chemicals and PP1 from Cayman Chemical.

Techniques: Infection, Isolation, Quantitative RT-PCR, Luciferase

Journal: bioRxiv

Article Title: SIRPA suppresses integrin-dependent virus endocytosis

doi: 10.64898/2026.04.17.719277

Figure Lengend Snippet: A) Differentiation of THP1 macrophages to M1 or M2 phenotypes increase SIRPA expression and decreases JUNV C1 infection. Student’s T test was used to determine significance. ***, P ≤ 0.05; **, P ≤ 0.002. B) Knockdown of SHP2 in M0 THP1 cells increases infection. Student’s T test with Welch’s correction was used to determine significance. ***, P ≤ 0.0005. C) BTT-3033 decreases JUNV C1 and TCRV infection in M0 THP1 cells. Student’s T test with Welch’s correction was used to determine significance. **, P ≤ 0.007; ***, P ≤ 0.0007. D) BTT-3033 decreases infection in M2 THP1 cells SIRPA knockdown cells. Left – western blots showing SIRPA expression in 5 different clones. Clone 5 was used for the infection experiment in the right panel. SIRPA surface expression for #5 is shown in Suppl. Fig. 6B. Two-way ANOVA was used to determine significance. *, P ≤ 0.02; **, P ≤ 0.002. E) PYK2 is more highly phosphorylated after incubation with TCRV or LCMV for 15 min. F) Knockdown of PYK2 in M0 THP1 cells causes lower levels of JUNV C1 internalization. Student’s T test was used to determine significance. ***, P ≤ 0.007.

Article Snippet: BTT-3033 and FAK inhibitor 14 (FIB-14) were purchased from TOCRIS, LHF-535 and favipiravir (6-fluoro-3-hydroxy-2-pyrazinecarboxamide) from TargetMol Chemicals and PP1 from Cayman Chemical.

Techniques: Expressing, Infection, Knockdown, Western Blot, Clone Assay, Incubation

Journal: bioRxiv

Article Title: Oncogenic RAS-driven α2 integrin induction under nutrient stress promotes cancer cell motility

doi: 10.64898/2026.04.02.716145

Figure Lengend Snippet: α 2 integrin was required for ECM uptake and ECM-dependent cell growth under amino acid starvation. (A,B) MCF10CA1 (A) and SW1990 (B) cells were grown in complete media or amino acid-free media for 18hr and seeded on 1 mg/ml collagen I labelled with pH-rodo (red), in complete media (Com) or amino acid-free media (AA). Cells were stained with Hoechst 33342 (blue) and imaged live with a Nikon A1 confocal microscope, 60x magnification. White shapes highlight the cell edges. Scale bar 30 μm. Collagen I uptake index was calculated with Image J. N=3 independent experiments. ****p < 0.0001 Mann-Whitney test. (C) MCF10CA1 cells were grown in amino acid-free media (AA) for 18 hours and seeded in AA-free media for 6 hours on 1 mg/ml collagen I labelled with pH-rodo (red). DMSO or 15 μM BTT-3033 (BTT) were added 2 hours after seeding. Cells were stained with Hoechst 33342 (blue) and imaged live with a Nikon A1 confocal microscope, 60x magnification. White shapes highlight the cell edges. Scale bar 30 μm. Collagen I uptake index was calculated with Image J. N=3 independent experiments. ****p < 0.0001 Mann-Whitney test. (D) MCF10CA1 cells were seeded on 2 mg/ml collagen I in amino acid-free media (AA) for 4 days in the presence of DMSO (control) or 15 μM BTT-3033 (BTT). Cells were fixed, stained with Hoechst 33342 and imaged with an ImageXpress micro. Data were analysed with MetaXpress. N=3 independent experiments, graph shows mean ± SEM. **p < 0.01 Kruskal-Wallis, Dunn’s multiple comparisons test. (E) SW1990 cells were seeded on 2 mg/ml collagen I in amino acid-free media (AA) for 7 days in the presence of DMSO (control) or 10 μM BTT-3033. Cells were fixed, stained and imaged as in D . N=3 independent experiments, graph shows mean ± SEM. *p < 0.05 Kruskal-Wallis, Dunn’s multiple comparisons test. (F) MCF10CA1 cells were seeded on 2 mg/ml collagen I, transfected with an siRNA targeting α2 integrin (Si-ITGA2) or a non- targeting siRNA (Si-nt) as a control and grown for 4 days in amino acid-free media (AA). Cells were fixed, stained and imaged as in D . N=5 independent experiments, graph shows mean ± SEM. **p < 0.01, ****p < 0.0001 Kruskal-Wallis, Dunn’s multiple comparisons test. (G) MCF10CA1 cells were seeded on plastic, transfected with an siRNA targeting α2 integrin (si-ITGA2) or a non-targeting siRNA (si-nt) as a control. Cell lysates were collected after 72 hours, samples were run by Western Blotting, membranes were stained for α2 integrin (ITGA2) and GAPDH and imaged with a Licor Odyssey Sa system. The protein band intensity was measured with Image Studio Lite software. N=4 independent experiments, graph shows mean ± SEM. *p =0.0286 Mann-Whitney test. Pink and purple ribbons are from wikicommons, http://creativecommons.org/licenses/by-sa/3.0/ .

Article Snippet: DMEM, high glucose, pyruvate; DMEM/F-12, glutamax; dialyzed foetal bovine serum (FBS), horse serum (HS) were from Gibco; DMEM, no amino acids from US Biological life sciences; FBS, Hydrocortisone and Insulin solution human from Sigma-Aldrich; Penicillin/ Streptomycin (Pen/strep) from Life technologies; Rat tail high concentration Collagen type I from Corning; Growth factor-reduced Matrigel from SLS; Hoechst 33342 from Invitrogen; NHS-Fluorescein, Phalloidin Alexa Fluor 555, Paraformaldehyde and pHrodo TM iFL Red STP ester from ThermoFisher; Vectashield mounting reagent with DAPI from VECTOR laboratories; 4-15% Mini PROTEAN TGX stain Free Protein gels from BioRad Laboratories; Protein ladder “Color Prestained Protein Standard” from New England Biolabs; High-Capacity cDNA Reverse Transcription kit from Applied biosystems; qPCRBIO SyGreen Blue Mix Lo-ROX from PCR BIOSYSTEMS; FITC-Anti Human CD49b (ITGA2) Antibody from Biolegend (#359306); Mouse anti-human CD49b (ITGA2) antibody from BD biosciences (#611017); P44/42 MAPK (ERK1/2) antibody (#9102) and Phospho-p44/42 MAPK (ERK1/2) antibody (#9101) from Cell signalling; GAPDH antibody from Santa Cruz Biotechnology (#sc-47724); α-Tubulin was from Cell signalling (#3873); IR Dye 680LT anti-Rabbit antibody and IR Dye 800LT anti-Mouse antibody from LICOR Biosciences; E64d (Aloxistatin) from AdooQ Bioscience; BTT-3033 and Torin-1 from Tocris Biosciences; MRTX1133, GCN2iB and RMC-6236 from MedChemExpress; Selumetinib from Selleck Chemicals.

Techniques: Staining, Microscopy, MANN-WHITNEY, Control, Transfection, Western Blot, Software

Journal: bioRxiv

Article Title: SIRPA suppresses integrin-dependent virus endocytosis

doi: 10.64898/2026.04.17.719277

Figure Lengend Snippet: A) shSIRPA or shCTRL cells were treated with BTT-3033 and infected with the indicated viruses. RNA was isolated at 24 hpi and viral RNA was measured by RT-qPCR. Shown is the average ± SD of 3–4 independent experiments. One-way ANOVA was used to determine significance. *, P ≤ 0.05; ***, P ≤ 0.001; ****, P ≤ 0.0001. B) VIAs were carried out with U2OS cells treated with BTT-3033. Internalized viral RNA was analyzed by RT-qPCR. Shown is the average ± SD of 3 independent experiments. Student’s T test was used to determine significance. **, P ≤ 0.01; ****, P ≤ 0.0001. C) shSIRPA or shCTRL cells were treated with BTT-3033 and infected with JUNV C1. Viral titers were analyzed at 24 hpi. Shown is the average ± SD of 3 independent experiments. One-way ANOVA was used to determine significance. ns, not significant, **, P ≤ 0.01; ****, P ≤ 0.0001. D) U2OS cells were treated with BTT-3033 at indicated concentrations and infected with MACV or VSV pseudoviruses. Luciferase assays were performed at 48 hpi. Shown is the average ± SD of 3 independent experiments. One-way ANOVA was used to determine significance. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. E) Viral titers in serum and spleen of BTT-3033– and favipravir-treated mice. Horizontal lines represent the mean for each group. The dotted horizontal lines represent the assay’s lower limit of detection. * , P ≤ 0.05; ** , P ≤ 0.01; *** , P ≤ 0.001 compared between groups. One animal in the DMSO placebo group was excluded from the analysis due to complications unrelated to the viral infection.

Article Snippet: BTT-3033 and FAK inhibitor 14 (FIB-14) were purchased from TOCRIS, LHF-535 and favipiravir (6-fluoro-3-hydroxy-2-pyrazinecarboxamide) from TargetMol Chemicals and PP1 from Cayman Chemical.

Techniques: Infection, Isolation, Quantitative RT-PCR, Luciferase

Journal: bioRxiv

Article Title: SIRPA suppresses integrin-dependent virus endocytosis

doi: 10.64898/2026.04.17.719277

Figure Lengend Snippet: A) Differentiation of THP1 macrophages to M1 or M2 phenotypes increase SIRPA expression and decreases JUNV C1 infection. Student’s T test was used to determine significance. ***, P ≤ 0.05; **, P ≤ 0.002. B) Knockdown of SHP2 in M0 THP1 cells increases infection. Student’s T test with Welch’s correction was used to determine significance. ***, P ≤ 0.0005. C) BTT-3033 decreases JUNV C1 and TCRV infection in M0 THP1 cells. Student’s T test with Welch’s correction was used to determine significance. **, P ≤ 0.007; ***, P ≤ 0.0007. D) BTT-3033 decreases infection in M2 THP1 cells SIRPA knockdown cells. Left – western blots showing SIRPA expression in 5 different clones. Clone 5 was used for the infection experiment in the right panel. SIRPA surface expression for #5 is shown in Suppl. Fig. 6B. Two-way ANOVA was used to determine significance. *, P ≤ 0.02; **, P ≤ 0.002. E) PYK2 is more highly phosphorylated after incubation with TCRV or LCMV for 15 min. F) Knockdown of PYK2 in M0 THP1 cells causes lower levels of JUNV C1 internalization. Student’s T test was used to determine significance. ***, P ≤ 0.007.

Article Snippet: BTT-3033 and FAK inhibitor 14 (FIB-14) were purchased from TOCRIS, LHF-535 and favipiravir (6-fluoro-3-hydroxy-2-pyrazinecarboxamide) from TargetMol Chemicals and PP1 from Cayman Chemical.

Techniques: Expressing, Infection, Knockdown, Western Blot, Clone Assay, Incubation

Journal: International Dental Journal

Article Title: NLRP12 Alleviated Periodontal Destruction via Suppressing Nuclear Factor Kappa-B Signalling Pathway

doi: 10.1016/j.identj.2026.109417

Figure Lengend Snippet: LPS inhibited the expression of NLRP12 and promoted the activation of the NF-κB signalling pathway in PDLSCs. (A) The expression level of NLRP12 in PDLSCs after LPS treatment was detected by immunofluorescence staining (scale bar = 50 μm). The qRT-PCR (B) and WB (C) analysis revealed significantly reduced expression of NLRP12 in PDLSCs after treatment with 10 µg/mL LPS. (D) WB analysis demonstrated that the expression of p-p65 and p-IκBα was significantly elevated relative to GAPDH in PDLSCs treated with 10 µg/mL LPS. Data were presented as mean ± SD ( n = 3). *** P < .001, **** P < .0001.

Article Snippet: After blocking with rapid sealing fluid for 30 minutes, the membranes were incubated with primary antibodies, including COL1 (#12256; CST), RUNX2 (#8486; CST), NLRP12 (#DF14960; Affinity Biosciences), phospho-p65 (p-p65) (#3033; CST), p65 (#8242; CST), p-IκBα (#2859; CST), IκBα (#4814; CST), GAPDH (#60004; Proteintech) overnight at 4°C.

Techniques: Expressing, Activation Assay, Immunofluorescence, Staining, Quantitative RT-PCR

Journal: International Dental Journal

Article Title: NLRP12 Alleviated Periodontal Destruction via Suppressing Nuclear Factor Kappa-B Signalling Pathway

doi: 10.1016/j.identj.2026.109417

Figure Lengend Snippet: Overexpression of NLRP12 alleviated the inflammatory responses and osteogenic differentiation inhibition of PDLSCs by suppressing the NF-κB pathway. (A) WB results showing the changes of protein expression levels of p-p65, p65, p-IκBα, and IκBα in PDLSCs after NLRP12 overexpression. (B) WB results showing the changes of protein expression levels of p-p65 and p65 in PDLSCs overexpressing NLRP12 after PMA treatment. (C)The qRT-PCR results demonstrating the transcriptional expression levels of IL-6, IL-8 , and IL-10 in PDLSCs overexpressing NLRP12 after PMA treatment. (D)WB results demonstrating alterations in the protein expression levels of COL1 and RUNX2 in PDLSCs overexpressing NLRP12 after PMA treatment. (E) Representative pictures showing ALP staining (scale bar = 500 μm) (F) Representative pictures showing alizarin red staining (scale bar = 500 μm). oeNC: PDLSCs transfected via negative control lentiviral. oeNLRP12: PDLSCs transfected via lentiviral with overexpression- NLRP12 . oeNC + LPS: PDLSCs transfected via negative control lentiviral and subsequently cultured under 10 µg/mL LPS. oeNLRP12 + LPS: PDLSCs transfected via lentiviral with overexpression- NLRP12 and subsequently cultured under 10 µg/mL LPS. Data were presented as mean ± SD ( n = 3). ns, no significant difference, * P < .05, ** P < .01, *** P < .001, **** P < .0001.

Article Snippet: After blocking with rapid sealing fluid for 30 minutes, the membranes were incubated with primary antibodies, including COL1 (#12256; CST), RUNX2 (#8486; CST), NLRP12 (#DF14960; Affinity Biosciences), phospho-p65 (p-p65) (#3033; CST), p65 (#8242; CST), p-IκBα (#2859; CST), IκBα (#4814; CST), GAPDH (#60004; Proteintech) overnight at 4°C.

Techniques: Over Expression, Inhibition, Expressing, Quantitative RT-PCR, Staining, Transfection, Negative Control, Cell Culture