mcherry e5d8f rabbit mab  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: ASFV early protein p30 suppresses antiviral type I IFN induction by targeting TRIM21 and RIG-I like receptor signaling adaptor MAVS

doi: 10.64898/2026.03.26.714469

Figure Lengend Snippet: Interactions exist between ASFV p30, TRIM21 and MAVS. ( A ) 293T cells were co-transfected with GFP-MAVS and Flag-TRIM21 for 24 h. Immunoprecipitation was performed using the rabbit anti-GFP mAb to confirm the interaction between exogenous MAVS and TRIM21. ( B ) 293T cells were co-transfected with mCherry-MAVS, Flag-TRIM21, and p30-GFP for 24 h. Immunoprecipitation was performed using rabbit mCherry pAb to confirm the interactions between exogenous MAVS, TRIM21 and p30. ( C ) MA104 cells were infected with ASFV for 72 h. Immunoprecipitation were performed using rabbit anti-MAVS mAb to confirm the interactions between endogenous MAVS, TRIM21 and p30. ( D ) The purified p30, TRIM21 and MAVS recombinant proteins were confirmed by SDS-PAGE and labeled with red boxes. ( E-G ) The interactions between purified proteins p30 and TRIM21, p30 and MAVS, and MAVS and TRIM21 verified through immunoprecipitations.

Article Snippet: The rabbit anti-TRIM21 mAb, mouse anti-TRIM21 mAb, rabbit anti-MAVS mAb, rabbit anti-IRF3 mAb, rabbit anti-Flag mAb, rabbit anti-HA mAb and rabbit mCherry pAb were all from ProteinTech (Wuhan, China).

Techniques: Transfection, Immunoprecipitation, Infection, Purification, Recombinant, SDS Page, Labeling

Journal: bioRxiv

Article Title: ASFV early protein p30 suppresses antiviral type I IFN induction by targeting TRIM21 and RIG-I like receptor signaling adaptor MAVS

doi: 10.64898/2026.03.26.714469

Figure Lengend Snippet: ASFV p30 inhibits TRIM21 upregulated MAVS signaling. ( A ) 293T cells were co-transfected mCherry-MAVS, Flag-TRIM21 and p30-GFP, as indicated, for 24 h. The downstream ISG56 and IFN-β mRNA levels activated by MAVS in the presence of TRIM21 were analyzed using RT-qPCR. ( B ) 3D4/21 cells were co-transfected with Flag-TRIM21 and p30-GFP, as indicated, for 12 h, and stimulated by transfection of poly (I:C) for 12 h. The downstream ISG56 and IFN-β mRNA levels activated by poly (I:C) in the presence of TRIM21 were analyzed using RT-qPCR. ( C ) 293T cells were co-transfected with mCherry-MAVS, Flag-TRIM21 and p30-GFP, plus ISRE Fluc and Rluc reporters, as indicated, for 24 h, and ISRE promoter activity was measured via dual luciferase reporter assay. ( D and E ) 3D4/21 cells were co-transfected with Flag-TRIM21 and p30-GFP for 12 h, as indicated. Cells were infected with VSV (D) or stimulated with transfection of poly (I:C) (E), followed by Western blotting for detection of cell signaling activation. ( F ) PAMs were transfected with p30 siRNA or control siRNA for 24 h, stimulated with transfection of poly (I:C) for 6 h, and then infected with ASFV for 48 h. The cell signaling and ASFV replication were detected by Western blotting.

Article Snippet: The rabbit anti-TRIM21 mAb, mouse anti-TRIM21 mAb, rabbit anti-MAVS mAb, rabbit anti-IRF3 mAb, rabbit anti-Flag mAb, rabbit anti-HA mAb and rabbit mCherry pAb were all from ProteinTech (Wuhan, China).

Techniques: Transfection, Quantitative RT-PCR, Activity Assay, Luciferase, Reporter Assay, Infection, Western Blot, Activation Assay, Control

Journal: bioRxiv

Article Title: ASFV early protein p30 suppresses antiviral type I IFN induction by targeting TRIM21 and RIG-I like receptor signaling adaptor MAVS

doi: 10.64898/2026.03.26.714469

Figure Lengend Snippet: ASFV p30 inhibits TRIM21 mediated ubiquitination of MAVS K27 ubiquitination. ( A ) 293T cells were co-transfected with Flag-TRIM21, mCherry-MAVS and HA-Ub for 24 h and then treated with MG-132 (10 μm) for 6 h. ( B ) 293T cells were co-transfected with Flag-TRIM21, mCherry-MAVS, p30-GFP, and HA-Ub for 24 h and then treated with MG-132 (10 μm) for 6 h. Cell samples were collected and immunoprecipitation were used for analysis of MAVS ubiquitination. ( C ) 293T cells were co-transfected with Flag-TRIM21, mCherry-MAVS and HA-Ub-K27 for 24 h and then treated with MG-132 (10 μm) for 6 h. ( D ) 293T cells were co-transfected with Flag-TRIM21, mCherry-MAVS, p30-myc, and HA-Ub-K27 for 24 h and then treated with MG-132 (10 μm) for 6 h. Cell samples were collected and immunoprecipitation were used for analysis of MAVS K27 ubiquitination.

Article Snippet: The rabbit anti-TRIM21 mAb, mouse anti-TRIM21 mAb, rabbit anti-MAVS mAb, rabbit anti-IRF3 mAb, rabbit anti-Flag mAb, rabbit anti-HA mAb and rabbit mCherry pAb were all from ProteinTech (Wuhan, China).

Techniques: Ubiquitin Proteomics, Transfection, Immunoprecipitation

Journal: bioRxiv

Article Title: Polycystin-1 C-Terminus Regulates Protein Synthesis-Related Pathways in Cardiomyocytes

doi: 10.64898/2026.03.21.713243

Figure Lengend Snippet: A . Maximum z-projected images from immunofluorescence confirm endogenous PC1 C-terminal localization with a striated pattern that resembles cardiomyocyte sarcomeres (white, 161F antibody) in hiPSC-CMs. siRNA-mediated PKD1 knockdown demonstrates antibody specificity. DAPI (blue) is used to label nuclei in all panels except panel D-E . Panels B–F are single confocal optical sections. The far-right panels show the raw fluorescence intensity profile for panel A and the normalized fluorescence intensity profiles for panels B, C , and F . B . Endogenous PC1 C-terminus (magenta) localizes in a striated pattern alternating with α-actinin staining (green), consistent with sarcomeric alignment. C . Endogenous PC1 C-terminus (magenta) shows minimal colocalization with the sarco-endoplasmic reticulum marker (KDEL, green). D, E . Representative live-cell images of hiPSC-CMs and adult mouse ventricular CMs expressing viral constructs encoding mCherry or mCherry-PC1-CT. PC1-CT localizes with a striated pattern and, in some cells, nuclear localization (zoom panels were renormalized to highlight localization). F . Localization of mCh-PC1-CT in adult vCMs aligns with Z-line proteins α-actinin (upper panel) and desmin (lower panel).

Article Snippet: Primary antibodies mouse anti-human PC1-CT (161F, 1:200) [ ], rabbit anti-α-actinin-2 (1:200, Invitrogen, #701914), rabbit anti-mCherry antibody (1:200, Cell Signaling, #43590), mouse anti-α-actinin2 (1:200, Invitrogen, #MAI-22863), mouse anti-desmin (1:100, Invitrogen, #MAS-13259), and rabbit anti-KDEL (1:100, Invitrogen, #PA1-013).

Techniques: Immunofluorescence, Knockdown, Fluorescence, Staining, Marker, Expressing, Construct