Journal: bioRxiv

Article Title: Phage display-mediated immuno-PCR to detect low-abundance secreted proteins in Drosophila

doi: 10.1101/2025.10.31.685841

Figure Lengend Snippet: ImpL2 nanobody screening using phage-displayed nanobody library. (A) Schematic of ImpL2 nanobody screening. A Maxisorp plate coated with mCherry-hIgG was used for negative selection, and a coated plate with ImpL2-hIgG was used for positive selection. Three iterative rounds of selection were conducted. (B) ELISA results using polyclonal phages from three rounds of selection. The coating amount of mCherry-hIgG and ImpL2-hIgG decreased across the rows. The intensity of the green color indicates the strength of the ELISA signal. (C) ELISA results of monoclonal phages isolated after the third round of selection. 96 individual clones were tested against the control protein (mCherry-hIgG) and the target protein (ImpL2-hIgG). (D) Immunostaining of cells expressing membrane-tethered ImpL2 using nanobody-displaying phages. GFP signals mark cells expressing membrane-tethered ImpL2. Bound phages were detected using an antibody against the M13 major coat protein. Nb.b201-displaying phage was used as a negative control. BF, bright field. (E) Immunostaining of cells expressing membrane-tethered ImpL2 using recombinant nanobodies. GFP signals mark cells expressing membrane-tethered ImpL2. Bound nanobodies were detected using an antibody against alpaca VHH fragments. Recombinant Nb127D01 was used as a negative control. BF, bright field.

Article Snippet: Other antibodies used in this study include anti-M13 major coat protein antibody (Santa Cruz Biotechnology, sc-53004 HRP and sc-53004 AF647), anti-alpaca VHH domain antibody (Jackson ImmunoResearch, 128-605-230 and 128-035-230) and anti-Human IgG Fc antibody (Thermo Fisher Scientific, A18829).

Techniques: Selection, Enzyme-linked Immunosorbent Assay, Isolation, Clone Assay, Control, Immunostaining, Expressing, Membrane, Negative Control, Recombinant

Journal: Cell

Article Title: SARS-CoV-2 mRNA vaccination induces functionally diverse antibodies to NTD, RBD, and S2

doi: 10.1016/j.cell.2021.06.005

Figure Lengend Snippet:

Article Snippet: M13 Reverse (−27) , Integrated DNA Technologies , 51-01-13-03.

Techniques: Virus, Recombinant, Lysis, Reverse Transcription, Amplification, Variant Assay, Nested PCR, Cloning, Plasmid Preparation, Software

Journal: Molecular oncology

Article Title: RING finger protein TOPORS modulates the expression of tumor suppressor SMAR1 in colorectal cancer via the TLR4-TRIF pathway.

doi: 10.1002/1878-0261.13126

Figure Lengend Snippet: Fig. 4. SMAR1-mediated regulation of STAT3 upon LPS treatment. Western blot showing reduced STAT3 expression in (A) CT26 cell line upon LPS treatment. SMAR1 and TOPORS expressions increase in CT26 (mouse colon cancer) cell line as well. (B) Representative FACS plot along with graph showing median fluorescent intensity. (C) Immunofluorescence images depicting the expression of SMAR1 and TOPORS in CT26 cell line. (D) Confocal images showing the expression of IRF3 and STAT1 under LPS-treated conditions in HT29 and CT26 cell lines. The scale bar represents 20 lm. Western blot representing the (E) expression status of pSTAT3 during SMAR1 knockout conditions in peritoneal macrophages (n = 6) with or without LPS treatment and (F) expression of SMAR1, TOPORS, and STATs in peritoneal macrophages during dose-dependent LPS treatment. (G) Representative FACS plots of M1 (F4/80+ CD 86+)- and M2 (F4/80+ CD 206+)-polarized peritoneal macrophage (n = 12) using CM collected from CT26 cells with or without LPS. b-Actin was used as an endogenous loading control in all the western blots. The values below the blot represent the fold change relative to control, which was calculated after normalization with b-actin. Data shown are representative of three independent experiments. Error bars indicate that all the values are mean SD, where *P < 0.05, **P < 0.01, ad ***P < 0.001 (two-way ANOVA, Bonferroni post-tests).

Article Snippet: The primary antibodies used for immunoblotting were SMAR1 (Bethyl Laboratories, Montgomery, TX, USA); b-actin and TLR4 (Santa Cruz Biotechnology); STAT1, STAT3, pSTAT1, pSTAT3, p-p65, p65, pJNK, JNK, IRF3, p-IRF3, and FLAG (Cell Signaling Technologies, Beverly, MA, USA); and TOPORS (Santa Cruz Biotechnology and Novus).

Techniques: Western Blot, Expressing, Knock-Out, Control

Journal: Molecular oncology

Article Title: RING finger protein TOPORS modulates the expression of tumor suppressor SMAR1 in colorectal cancer via the TLR4-TRIF pathway.

doi: 10.1002/1878-0261.13126

Figure Lengend Snippet: Fig. 6. Working model showing the regulation of SMAR1 upon LPS treatment. TLR4 upon recognizing LPS initiates the signaling cascade. This leads to the activation of transcription factor IRF3 via TLR4-TRIF-dependent pathway. IRF3 upon phosphorylation translocates to the nucleus, where it regulates the expression of various type I interferons including IFN-b. It could also induce the expression of zinc finger protein TOPORS, which further controls the transcription of tumor suppressor SMAR1 via binding to the consensus region present on its promoter. LPS-induced SMAR1 in return represses the transcription of STAT3 and hence shifts the TAMs toward antitumor M1 profile, whereas upon inhibiting TLR4 pathway using a chemical inhibitor (TAK-242), the downstream adapter proteins fail to bind to the TIR domain of TLR4. Under such circumstances, even LPS stimulation fails to enhance the expression of TOPORS and SMAR1. In the absence of effective SMAR1 concentration, the repression on STAT3 promoter is released. In excess STAT3 conditions, most of the TAMs tend to polarize to M2 phenotype, which is associated with cancer growth and progression.

Article Snippet: The primary antibodies used for immunoblotting were SMAR1 (Bethyl Laboratories, Montgomery, TX, USA); b-actin and TLR4 (Santa Cruz Biotechnology); STAT1, STAT3, pSTAT1, pSTAT3, p-p65, p65, pJNK, JNK, IRF3, p-IRF3, and FLAG (Cell Signaling Technologies, Beverly, MA, USA); and TOPORS (Santa Cruz Biotechnology and Novus).

Techniques: Activation Assay, Phospho-proteomics, Expressing, Binding Assay, Concentration Assay