shp 1 inhibitor  (MedChemExpress)

Journal: bioRxiv

Article Title: SHP-1 interacts with NFκB1 to inhibit its phosphorylation and nuclear translocation to suppress excessive bacterial inflammation

doi: 10.1101/2024.04.13.589348

Figure Lengend Snippet: ( A ) Schematic diagram of individual experiments on Chinese tongue sole. CO, blank control group; NC, PBS injection followed by V. anguillarum infection; OE, SHP-1 overexpression followed by V. anguillarum ; IN, SHP-1 inhibition followed by V. anguillarum infection. ( B ) Morphological changes in the intestine, liver, spleen, and gill of Chinese tongue sole after infection with V. anguillarum . ( C ) Survival of Chinese tongue sole after infection with V. anguillarum at LD80. N = 60 individuals in each group. ( D ) Survival of Chinese tongue sole after infection with V. anguillarum at LD50. N = 60 individuals in each group. ( E ) Histopathological images of liver (upper three rows) and intestine (lower three rows) of Chinese tongue sole at 0 h, 12 h, 24 h, 48 h, 72 h after V. anguillarum infection at LD80. The long arrow pointing to the left represents inflammatory cells infiltration; The short arrow pointing to the left represents vacuolar degeneration of liver cells; The long arrow pointing to the right represents separation of intestinal mucosal layer and submucosal layer, intestinal villus rupture. Scale bars, 100 μm (E).

Article Snippet: The SHP-1 inhibitor named TPI (Topscience, China) was added to hepatocytes and primary macrophages to reach 5 μmol in a total volume of 2 mL L15 medium and incubated for 24 h.

Techniques: Control, Injection, Infection, Over Expression, Inhibition

Journal: bioRxiv

Article Title: SHP-1 interacts with NFκB1 to inhibit its phosphorylation and nuclear translocation to suppress excessive bacterial inflammation

doi: 10.1101/2024.04.13.589348

Figure Lengend Snippet: ( A to H ) Expression of il-1β , il-6, tlr5 , myd88 , nfκb1 , nfκb2 , jak2 , stat3 in the liver of Chinese tongue sole after infection with V. anguillarum at LD80. ( I ) Expression of SHP-1, IL-1β, IL-6 of liver tissue by tyramide signal amplification technology. CO, blank control group; NC, PBS injection followed by V. anguillarum infection; OE, SHP-1 overexpression followed by V. anguillarum ; IN, SHP-1 inhibition followed by V. anguillarum infection. DAPI staining was used to indicate the nuclei with blue fluorescence; SHP-1 combined with CoraLite488-conjugated goat anti-rabbit IgG secondary antibody shows green fluorescence, IL-1β combined with Cy3 goat anti- rabbit IgG secondary antibody shows pink fluorescence; IL-6 combined with Cy5 goat anti-rabbit IgG secondary antibody displays red fluorescence. Data are presented as means ± SD, n = 3 biologically independent experiments [(A) to (H)]. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by one-way ANOVA. Scale bars, 50 μm (I).

Article Snippet: The SHP-1 inhibitor named TPI (Topscience, China) was added to hepatocytes and primary macrophages to reach 5 μmol in a total volume of 2 mL L15 medium and incubated for 24 h.

Techniques: Expressing, Infection, Amplification, Control, Injection, Over Expression, Inhibition, Staining, Fluorescence

Journal: bioRxiv

Article Title: SHP-1 interacts with NFκB1 to inhibit its phosphorylation and nuclear translocation to suppress excessive bacterial inflammation

doi: 10.1101/2024.04.13.589348

Figure Lengend Snippet: ( A to C ) Expression of SHP-1 in hepatocytes after LPS stimulation. ( D ) Expression of il-1β , il-6 , tlr5 , myd88 , nfκb1 , nfκb2 , jak2 , and stat3 in hepatocytes treated or untreated with LPS. ( E to G ) Expression of SHP-1 in macrophages after LPS stimulation. ( H ) Expression of il-1β , il-6 , tlr5 , myd88 , nfκb1 , nfκb2 , jak2 , and stat3 in macrophages treated or untreated with LPS. ( I ) Macrophage phagocytosis determined by measuring the number of macrophages that engulfed fluorescent microspheres via immunofluorescence experiments. DAPI is used to stain the cell nucleus as blue fluorescence. SHP-1 displays green fluorescence, which is located in the nucleus and cytoplasm of macrophages, by specifically binding to the CoraLite488-conjugated goat anti-rabbit IgG secondary antibody; The fluorescent microspheres can be excited by a wavelength of 565 nm to produce red fluorescence. ( J ) Phagocytosis measured by the absorbance of neutral red engulfed by macrophages. ( K ) Macrophage polarization markers including nos1 , arg-2 , and il-10 were determined by qRT-PCR. Data are presented as means ± SD, n = 3 biologically independent experiments [(A) to (K)]. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by t test. Scale bars, 50 μm [(I) and (J)].

Article Snippet: The SHP-1 inhibitor named TPI (Topscience, China) was added to hepatocytes and primary macrophages to reach 5 μmol in a total volume of 2 mL L15 medium and incubated for 24 h.

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence, Binding Assay, Quantitative RT-PCR

Journal: bioRxiv

Article Title: SHP-1 interacts with NFκB1 to inhibit its phosphorylation and nuclear translocation to suppress excessive bacterial inflammation

doi: 10.1101/2024.04.13.589348

Figure Lengend Snippet: ( A to H ) Overexpression of SHP-1 in hepatocytes and macrophages validated by fluorescence microscopy imaging, qRT-PCR, and Western blot. Plasmid indicates transfection of SHP-1 plasmid; PNC indicates transfection of empty vector as negative control. (I and J ) Expression of il-1β , il-6 , tlr5 , myd88 , nfκb1 , nfκb2 , jak2 , and stat3 in hepatocytes and macrophages which were transfected with SHP-1 plasmid or empty vector followed by LPS stimulation. ( K ) Macrophage phagocytosis determined by measuring the number of macrophages that engulfed fluorescent microspheres via immunofluorescence experiments. DAPI is used to stain the cell nucleus as blue fluorescence. SHP-1 displays green fluorescence, which is located in the nucleus and cytoplasm of macrophages, by specifically binding to the CoraLite488-conjugated goat anti-rabbit IgG secondary antibody; The fluorescent microspheres can be excited by a wavelength of 565 nm to produce red fluorescence. ( L ) Phagocytosis measured by the absorbance of neutral red engulfed by macrophages. ( M ) Macrophage polarization markers including nos1 , arg-2 , and il-10 were determined by qRT-PCR. Data are presented as means ± SD, n = 3 biologically independent experiments [(A) to (M)]. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by t test. Scale bars, 50 μm [(K) and (L)].

Article Snippet: The SHP-1 inhibitor named TPI (Topscience, China) was added to hepatocytes and primary macrophages to reach 5 μmol in a total volume of 2 mL L15 medium and incubated for 24 h.

Techniques: Over Expression, Fluorescence, Microscopy, Imaging, Quantitative RT-PCR, Western Blot, Plasmid Preparation, Transfection, Negative Control, Expressing, Immunofluorescence, Staining, Binding Assay

Journal: bioRxiv

Article Title: SHP-1 interacts with NFκB1 to inhibit its phosphorylation and nuclear translocation to suppress excessive bacterial inflammation

doi: 10.1101/2024.04.13.589348

Figure Lengend Snippet: ( A to H ) Silencing SHP-1 in hepatocytes and macrophages validated by fluorescence microscopy imaging, qRT-PCR, and Western blot. siRNA indicates transfection of shp-1 siRNA; SNC indicates transfection of randomly synthesized siRNA as negative control. ( I and J ) Expression of il-1β , il-6 , tlr5 , myd88 , nfκb1 , nfκb2 , jak2 , and stat3 in hepatocytes and macrophages which were transfected with shp-1 siRNA or randomly synthesized siRNA followed by LPS stimulation. (K ) Macrophage phagocytosis determined by measuring the number of macrophages that engulfed fluorescent microspheres via immunofluorescence experiments. DAPI is used to stain the cell nucleus as blue fluorescence. SHP-1 displays green fluorescence, which is located in the nucleus and cytoplasm of macrophages, by specifically binding to the CoraLite488-conjugated goat anti-rabbit IgG secondary antibody; The fluorescent microspheres can be excited by a wavelength of 565 nm to produce red fluorescence. ( L ) Phagocytosis measured by the absorbance of neutral red engulfed by macrophages. ( M ) Macrophage polarization markers including nos1 , arg-2 , and il-10 were determined by qRT-PCR. Data are presented as means ± SD, n = 3 biologically independent experiments [(A) to (M)]. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by t test. Scale bars, 50 μm [(K) and (L)].

Article Snippet: The SHP-1 inhibitor named TPI (Topscience, China) was added to hepatocytes and primary macrophages to reach 5 μmol in a total volume of 2 mL L15 medium and incubated for 24 h.

Techniques: Fluorescence, Microscopy, Imaging, Quantitative RT-PCR, Western Blot, Transfection, Synthesized, Negative Control, Expressing, Immunofluorescence, Staining, Binding Assay

Journal: bioRxiv

Article Title: SHP-1 interacts with NFκB1 to inhibit its phosphorylation and nuclear translocation to suppress excessive bacterial inflammation

doi: 10.1101/2024.04.13.589348

Figure Lengend Snippet: ( A to C ) SHP-1 inhibition in hepatocytes validated by qRT-PCR and Western blot. ( D ) Expression of il- 1β , myd88 , nfκb1 , nfκb2, il-6 , jak2 , stat3 , and tlr5 in hepatocytes which were incubated with SHP-1 inhibitor or PBS followed by LPS stimulation. ( E to G ) SHP-1 inhibition in macrophages validated by qRT-PCR and Western blot. Inhibitor indicated incubation with SHP-1 inhibitor; INC indicated incubation with PBS as negative control. ( H ) Expression of il-1β , myd88 , nfκb1 , nfκb2, il-6 , jak2 , stat3 , tlr5 in macrophages which were incubated with SHP-1 inhibitor or PBS followed by LPS stimulation. ( I ) Macrophage phagocytosis determined by measuring the number of macrophages that engulfed fluorescent microspheres via immunofluorescence experiments. DAPI is used to stain the cell nucleus as blue fluorescence. SHP-1 displays green fluorescence, which is located in the nucleus and cytoplasm of macrophages, by specifically binding to the CoraLite488-conjugated goat anti-rabbit IgG secondary antibody; The fluorescent microspheres can be excited by a wavelength of 565 nm to produce red fluorescence. (J) Phagocytosis measured by the absorbance of neutral red engulfed by macrophages. (K) Macrophage polarization markers including nos1 , arg-2 , and il-10 were determined by qRT-PCR. Data are presented as means ± SD, n = 3 biologically independent experiments [(A) to (K)]. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by t test. Scale bars, 50 μm [(I) and (J)].

Article Snippet: The SHP-1 inhibitor named TPI (Topscience, China) was added to hepatocytes and primary macrophages to reach 5 μmol in a total volume of 2 mL L15 medium and incubated for 24 h.

Techniques: Inhibition, Quantitative RT-PCR, Western Blot, Expressing, Incubation, Negative Control, Immunofluorescence, Staining, Fluorescence, Binding Assay

Journal: bioRxiv

Article Title: SHP-1 interacts with NFκB1 to inhibit its phosphorylation and nuclear translocation to suppress excessive bacterial inflammation

doi: 10.1101/2024.04.13.589348

Figure Lengend Snippet: ( A ) The respiratory burst activity, SOD and MDA in macrophages treated or untreated with LPS. ( B ) The respiratory burst activity, SOD and MDA in macrophages transfected with shp- 1 plasmid or empty vector group. ( C ) The respiratory burst activity, SOD and MDA in macrophages transfected with shp-1 siRNA or randomly synthesized siRNA. ( D ) The respiratory burst activity, SOD and MDA in macrophages treated with SHP-1 inhibitor or PBS. ( E to F ) The SOD and MDA level in the liver of Chinese tongue sole in different groups. CO, blank control group; NC, PBS injection followed by V. anguillarum infection; OE, SHP-1 overexpression followed by V. anguillarum ; IN, SHP-1 inhibition followed by V. anguillarum infection. Data are presented as means ± SD, n = 3 biologically independent experiments [(A) to (F)]. [(A) to (D)] * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by t test. [(E) to (F)] * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by one-way ANOVA.

Article Snippet: The SHP-1 inhibitor named TPI (Topscience, China) was added to hepatocytes and primary macrophages to reach 5 μmol in a total volume of 2 mL L15 medium and incubated for 24 h.

Techniques: Activity Assay, Transfection, Plasmid Preparation, Synthesized, Control, Injection, Infection, Over Expression, Inhibition

Journal: bioRxiv

Article Title: SHP-1 interacts with NFκB1 to inhibit its phosphorylation and nuclear translocation to suppress excessive bacterial inflammation

doi: 10.1101/2024.04.13.589348

Figure Lengend Snippet: ( A and B ) Expression of SHP-1, IL-1β, IL-6, TLR5, MYD88, NFκB1, pNFκB1 in the liver of Chinese tongue sole after the infection of V. anguillarum (LD80) by Western blot. CO, blank control group; NC, PBS injection followed by V. anguillarum infection; OE, SHP-1 overexpression followed by V. anguillarum ; IN, SHP-1 inhibition followed by V. anguillarum infection. ( C to E ) NFκB1 protein content in cytoplasm and nucleus detected by Western blot. The internal references are GAPDH and Lamin A/C respectively. ( F ) Immunofluorescence and immunohistochemistry analysis of NFκB1 of different groups. DAPI is used to stain the cell nuclei with blue fluorescence; The SHP-1 combined with CoraLite488- conjugated goat anti-rabbit IgG secondary antibody shows green fluorescence; NFκB1 combined with Cy5 goat anti-rabbit IgG secondary antibody displays red fluorescence; Brown represents a positive immunohistochemical result and cell nucleus was stained blue. White arrow represents NFκB1 in the nucleus. Data are presented as means ± SD, n = 3 biologically independent experiments [(A) to (E)]. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by one-way ANOVA. Scale bars, 50 μm (F).

Article Snippet: The SHP-1 inhibitor named TPI (Topscience, China) was added to hepatocytes and primary macrophages to reach 5 μmol in a total volume of 2 mL L15 medium and incubated for 24 h.

Techniques: Expressing, Infection, Western Blot, Control, Injection, Over Expression, Inhibition, Immunofluorescence, Immunohistochemistry, Staining, Fluorescence, Immunohistochemical staining

Journal: bioRxiv

Article Title: SHP-1 interacts with NFκB1 to inhibit its phosphorylation and nuclear translocation to suppress excessive bacterial inflammation

doi: 10.1101/2024.04.13.589348

Figure Lengend Snippet: ( A ) Co-immunoprecipitation experiment used to examine the interaction between SHP-1 and NFκB1. Co-IP was performed with Flag-labeled magnetic beads. ( B ) Immunofluorescence co-localization analysis of SHP-1 and NFκB1 in hepatocytes, blue fluorescence represents nucleus, green fluorescence represents SHP-1 proteins, red fluorescence represents NFκB1 proteins, Merge represents combining three fluorescent images together. DAPI is used to stain the cell nucleus as blue fluorescence; SHP-1 combined with CoraLite488-conjugated goat anti-rabbit IgG secondary antibody shows green fluorescence; NFκB1 combined with Cy5 goat anti-rabbit IgG secondary antibody shows red fluorescence; White arrow represents SHP-1 and NFκB1 co- localization. ( C ) Schematic diagram of SHP-1 regulating NFκB signaling pathway. Mechanisms revealed in this study are highlighted in red. Scale bars, 50 μm (B).

Article Snippet: The SHP-1 inhibitor named TPI (Topscience, China) was added to hepatocytes and primary macrophages to reach 5 μmol in a total volume of 2 mL L15 medium and incubated for 24 h.

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Labeling, Magnetic Beads, Immunofluorescence, Fluorescence, Staining