rabbit anti nfkb2 p52  (Proteintech)

Journal: bioRxiv

Article Title: A Suite of Eight Toxoplasma gondii Effectors Cooperates to Activate the Non-canonical NF-κB Pathway

doi: 10.64898/2026.03.12.711255

Figure Lengend Snippet: A) Top-scoring transcription factors predicted to regulate the MYR1-dependent upregulated genes in HFF cells relative to uninfected controls were identified by analysis of previously published data using Enrichr. B-C) Representative images (top) and quantitative analysis (bottom) for RelB (B) and p52 (C) nuclear accumulation in HFFs infected with RH (WT), Δ myr1 or Δ myr1 ::MYR1 complement parasites. Twenty-four hours post-infection, cells were fixed and labeled with DAPI (blue), anti-GAP45 (red), and either anti-RelB or anti-p52 (green). Scale bars = 10 µm. Plots display the nuclear-to-cytoplasmic signal ratios for at least 300 infected cells (red arrows) per condition; uninfected cells are indicated by white arrows. Data from three independent experiments were combined for analysis. The horizontal dashed lines indicate the mean. Statistical significance was determined using a one-way ANOVA with Tukey’s multiple comparison test; ****P < 0.0001, ns = not significant.

Article Snippet: Primary antibodies used were: rabbit anti-phospho-NF-kappaB2 p100 (Ser866/870) (1:1,000; Cell Signaling Technology, #4810), rabbit anti-NF-kappaB2 p100/p52 (1:1,000; Cell Signaling Technology, #4882), rabbit anti-NIK (1:1,000; Cell Signaling Technology, #4994), rabbit anti-TRAF3 (1:1,000; Cell Signaling Technology, #4729), and mouse anti-β-tubulin (Developmental Studies Hybridoma Bank, AB_528499), which detects both parasite and host β-tubulin and was used as a loading control.

Techniques: Infection, Labeling, Comparison

Journal: bioRxiv

Article Title: A Suite of Eight Toxoplasma gondii Effectors Cooperates to Activate the Non-canonical NF-κB Pathway

doi: 10.64898/2026.03.12.711255

Figure Lengend Snippet: A-B) Representative images (top) and quantitative analysis (bottom) for RelB (A) and p52 (B) nuclear accumulation in MEFs infected with RH (WT), Δ myr1 or Δ myr1 ::MYR1 complement parasites. Twenty-four hours post-infection, cells were fixed and labeled with DAPI (blue), anti-GAP45 (red), and either anti-RelB or anti-p52 (green). Scale bars = 10 µm. Plots display the nuclear-to-cytoplasmic signal ratios for at least 300 infected cells (red arrows) per condition; uninfected cells are indicated by white arrows. Data from three independent experiments were combined for analysis. The horizontal dashed lines indicate the mean. Statistical significance was determined using a one-way ANOVA with Tukey’s multiple comparison test; ****P < 0.0001, ns = not significant.

Article Snippet: Primary antibodies used were: rabbit anti-phospho-NF-kappaB2 p100 (Ser866/870) (1:1,000; Cell Signaling Technology, #4810), rabbit anti-NF-kappaB2 p100/p52 (1:1,000; Cell Signaling Technology, #4882), rabbit anti-NIK (1:1,000; Cell Signaling Technology, #4994), rabbit anti-TRAF3 (1:1,000; Cell Signaling Technology, #4729), and mouse anti-β-tubulin (Developmental Studies Hybridoma Bank, AB_528499), which detects both parasite and host β-tubulin and was used as a loading control.

Techniques: Infection, Labeling, Comparison

Journal: bioRxiv

Article Title: A Suite of Eight Toxoplasma gondii Effectors Cooperates to Activate the Non-canonical NF-κB Pathway

doi: 10.64898/2026.03.12.711255

Figure Lengend Snippet: T. gondii activates the non-canonical NF-κB pathway through MYR1-dependent TRAF3 depletion and NIK stabilization. (A) Representative images (top) and quantitative analysis (bottom) for RelB nuclear accumulation in HFFs infected with RH (WT) parasites over a 24-h time course. At 6, 12, 18, and 24 h post-infection, cells were fixed and labeled with DAPI (blue), anti-GAP45 (red), and anti-RelB (green). Scale bars = 10 µm. Plots display the nuclear-to-cytoplasmic intensity ratios for at least 300 infected cells (red arrows) per condition; uninfected cells are indicated by white arrows. Data from three independent experiments were combined for analysis. The horizontal dashed lines indicate the mean. Statistical significance was determined using a one-way ANOVA with Tukey’s multiple comparison test; ***P < 0.001, ****P < 0.0001, ns = not significant. (B) Immunoblot analysis (top) and corresponding quantification (bottom) of TRAF3, NIK, p100 phosphorylation, and p100-to-p52 processing in HFFs that were uninfected (UI) or infected with RH (WT), Δmyr1 or Δmyr1 ::MYR1 complement parasites. Lysates collected 24 h post-infection were resolved by SDS-PAGE and probed with specific primary antibodies; β-tubulin served as a loading control. Band intensities were measured using ImageJ and normalized to β-tubulin. Data are presented as mean ±SD from three biological replicates. Statistical significance for all panels was determined using a one-way ANOVA with Tukey’s multiple comparison test; **P < 0.01, ***P < 0.001, ****P < 0.0001, ns = not significant

Article Snippet: Primary antibodies used were: rabbit anti-phospho-NF-kappaB2 p100 (Ser866/870) (1:1,000; Cell Signaling Technology, #4810), rabbit anti-NF-kappaB2 p100/p52 (1:1,000; Cell Signaling Technology, #4882), rabbit anti-NIK (1:1,000; Cell Signaling Technology, #4994), rabbit anti-TRAF3 (1:1,000; Cell Signaling Technology, #4729), and mouse anti-β-tubulin (Developmental Studies Hybridoma Bank, AB_528499), which detects both parasite and host β-tubulin and was used as a loading control.

Techniques: Infection, Labeling, Comparison, Western Blot, Phospho-proteomics, SDS Page, Control

Journal: Cancer Biology & Medicine

Article Title: Migration and invasion inhibitory protein inhibits M2 macrophage polarization to suppress colorectal cancer progression through the STING–NFκB2–IL10 axis

doi: 10.20892/j.issn.2095-3941.2025.0282

Figure Lengend Snippet: MIIP modulates the STING–NFκB2–IL10 signaling pathway in CRC cells. Immunofluorescence staining showing the effects of MIIP knockdown and MIIP overexpression on dsDNA levels (A) and the distribution and expression of STING (B) in SW480 and SW620 cells. (C) Western blots showing the effects of MIIP knockdown and overexpression on STING, TRAF3, p-p100, and p52 levels. (D) ELISA results showing the effects of MIIP knockdown and overexpression on IL10 levels in the cell culture medium. (E and F) Effects of MIIP knockdown and/or STING knockdown on p52 expression (E) and IL10 levels in the cell culture medium (F). All data are presented as the means ± SDs ( n = 3). * P < 0.05, ** P < 0.01, and *** P < 0.001; scale bar, 50 μm.

Article Snippet: The primary antibodies used included mouse anti-dsDNA (1:1000, ab270732; Abcam, Cambridge, MA, USA), rabbit anti-NFKB2/p52 (1:200, 15503-1-AP; Proteintech, Wuhan, Hubei, China), rabbit anti-TMEM173/STING (1:200, 19851-1-AP; Proteintech, Wuhan, Hubei, China), and rabbit anti-IL-10 antibodies (1:100, 20850-1-AP; Proteintech, Wuhan, Hubei, China).

Techniques: Immunofluorescence, Staining, Knockdown, Over Expression, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Cancer Biology & Medicine

Article Title: Migration and invasion inhibitory protein inhibits M2 macrophage polarization to suppress colorectal cancer progression through the STING–NFκB2–IL10 axis

doi: 10.20892/j.issn.2095-3941.2025.0282

Figure Lengend Snippet: MIIP inhibits tumor growth and metastasis and M2 macrophage infiltration in a syngeneic CRC mouse model through the STING–NFκB2–IL10 axis. (A) Tumors in situ (indicated by green arrows) and liver metastatic nodules (indicated by black arrows) were observed in a syngeneic CRC mouse model. Statistics for the size of tumors in situ (B) and the number of metastatic foci (C). (D) Statistical analysis of IL10 levels in mouse blood. (E) Representative images of HE and IHC staining for SATB2, MIIP, STING, IL10, p52, and CD163 (green arrows) in tumor tissues from the different groups. The bar graphs show the percentages of high/low STING (F), IL-10 (G), and p52 (H) expression in the different groups. (I) Comparison of the number of CD163-positive cells among the different groups. All data are presented as the means ± SDs ( n = 3). ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001; scale bar, 50 μm.

Article Snippet: The primary antibodies used included mouse anti-dsDNA (1:1000, ab270732; Abcam, Cambridge, MA, USA), rabbit anti-NFKB2/p52 (1:200, 15503-1-AP; Proteintech, Wuhan, Hubei, China), rabbit anti-TMEM173/STING (1:200, 19851-1-AP; Proteintech, Wuhan, Hubei, China), and rabbit anti-IL-10 antibodies (1:100, 20850-1-AP; Proteintech, Wuhan, Hubei, China).

Techniques: In Situ, Immunohistochemistry, Expressing, Comparison

Journal: Cancer Biology & Medicine

Article Title: Migration and invasion inhibitory protein inhibits M2 macrophage polarization to suppress colorectal cancer progression through the STING–NFκB2–IL10 axis

doi: 10.20892/j.issn.2095-3941.2025.0282

Figure Lengend Snippet: MIIP modulates the STING–NFκB2–IL10 signaling pathway in CRC cells. Immunofluorescence staining showing the effects of MIIP knockdown and MIIP overexpression on dsDNA levels (A) and the distribution and expression of STING (B) in SW480 and SW620 cells. (C) Western blots showing the effects of MIIP knockdown and overexpression on STING, TRAF3, p-p100, and p52 levels. (D) ELISA results showing the effects of MIIP knockdown and overexpression on IL10 levels in the cell culture medium. (E and F) Effects of MIIP knockdown and/or STING knockdown on p52 expression (E) and IL10 levels in the cell culture medium (F). All data are presented as the means ± SDs ( n = 3). * P < 0.05, ** P < 0.01, and *** P < 0.001; scale bar, 50 μm.

Article Snippet: The primary antibodies used included rabbit anti-MIIP (1:1000, HPA044948; Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-MIIP (1:1000, orb537083; Biorbyt, Cambridge, UK), rabbit anti-STING (1:2000, 19851-1-AP; Proteintech, Wuhan, Hubei, China), mouse anti-TRAF3 (1:1000, 66310-1-Ig; Proteintech, Wuhan, Hubei, China), rabbit anti-p52 (1:1000, 4882S; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-p52 (1:500, 15503-1-AP; Proteintech, Wuhan, Hubei, China), rabbit anti-p-p100 (1:1000, 4810T; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-CD163 (1:1000, 68922; Cell Signaling Technology, Danvers, MA, USA), and mouse anti-β-actin antibodies (1:1000, 3700S; Cell Signaling Technology, Danvers, MA, USA).

Techniques: Immunofluorescence, Staining, Knockdown, Over Expression, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Cancer Biology & Medicine

Article Title: Migration and invasion inhibitory protein inhibits M2 macrophage polarization to suppress colorectal cancer progression through the STING–NFκB2–IL10 axis

doi: 10.20892/j.issn.2095-3941.2025.0282

Figure Lengend Snippet: MIIP inhibits tumor growth and metastasis and M2 macrophage infiltration in a syngeneic CRC mouse model through the STING–NFκB2–IL10 axis. (A) Tumors in situ (indicated by green arrows) and liver metastatic nodules (indicated by black arrows) were observed in a syngeneic CRC mouse model. Statistics for the size of tumors in situ (B) and the number of metastatic foci (C). (D) Statistical analysis of IL10 levels in mouse blood. (E) Representative images of HE and IHC staining for SATB2, MIIP, STING, IL10, p52, and CD163 (green arrows) in tumor tissues from the different groups. The bar graphs show the percentages of high/low STING (F), IL-10 (G), and p52 (H) expression in the different groups. (I) Comparison of the number of CD163-positive cells among the different groups. All data are presented as the means ± SDs ( n = 3). ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001; scale bar, 50 μm.

Article Snippet: The primary antibodies used included rabbit anti-MIIP (1:1000, HPA044948; Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-MIIP (1:1000, orb537083; Biorbyt, Cambridge, UK), rabbit anti-STING (1:2000, 19851-1-AP; Proteintech, Wuhan, Hubei, China), mouse anti-TRAF3 (1:1000, 66310-1-Ig; Proteintech, Wuhan, Hubei, China), rabbit anti-p52 (1:1000, 4882S; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-p52 (1:500, 15503-1-AP; Proteintech, Wuhan, Hubei, China), rabbit anti-p-p100 (1:1000, 4810T; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-CD163 (1:1000, 68922; Cell Signaling Technology, Danvers, MA, USA), and mouse anti-β-actin antibodies (1:1000, 3700S; Cell Signaling Technology, Danvers, MA, USA).

Techniques: In Situ, Immunohistochemistry, Expressing, Comparison

Journal: Cancer Biology & Medicine

Article Title: Migration and invasion inhibitory protein inhibits M2 macrophage polarization to suppress colorectal cancer progression through the STING–NFκB2–IL10 axis

doi: 10.20892/j.issn.2095-3941.2025.0282

Figure Lengend Snippet: MIIP modulates the STING–NFκB2–IL10 signaling pathway in CRC cells. Immunofluorescence staining showing the effects of MIIP knockdown and MIIP overexpression on dsDNA levels (A) and the distribution and expression of STING (B) in SW480 and SW620 cells. (C) Western blots showing the effects of MIIP knockdown and overexpression on STING, TRAF3, p-p100, and p52 levels. (D) ELISA results showing the effects of MIIP knockdown and overexpression on IL10 levels in the cell culture medium. (E and F) Effects of MIIP knockdown and/or STING knockdown on p52 expression (E) and IL10 levels in the cell culture medium (F). All data are presented as the means ± SDs ( n = 3). * P < 0.05, ** P < 0.01, and *** P < 0.001; scale bar, 50 μm.

Article Snippet: The primary antibodies used included rabbit anti-MIIP (1:1000, HPA044948; Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-MIIP (1:1000, orb537083; Biorbyt, Cambridge, UK), rabbit anti-STING (1:2000, 19851-1-AP; Proteintech, Wuhan, Hubei, China), mouse anti-TRAF3 (1:1000, 66310-1-Ig; Proteintech, Wuhan, Hubei, China), rabbit anti-p52 (1:1000, 4882S; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-p52 (1:500, 15503-1-AP; Proteintech, Wuhan, Hubei, China), rabbit anti-p-p100 (1:1000, 4810T; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-CD163 (1:1000, 68922; Cell Signaling Technology, Danvers, MA, USA), and mouse anti-β-actin antibodies (1:1000, 3700S; Cell Signaling Technology, Danvers, MA, USA).

Techniques: Immunofluorescence, Staining, Knockdown, Over Expression, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Cancer Biology & Medicine

Article Title: Migration and invasion inhibitory protein inhibits M2 macrophage polarization to suppress colorectal cancer progression through the STING–NFκB2–IL10 axis

doi: 10.20892/j.issn.2095-3941.2025.0282

Figure Lengend Snippet: MIIP inhibits tumor growth and metastasis and M2 macrophage infiltration in a syngeneic CRC mouse model through the STING–NFκB2–IL10 axis. (A) Tumors in situ (indicated by green arrows) and liver metastatic nodules (indicated by black arrows) were observed in a syngeneic CRC mouse model. Statistics for the size of tumors in situ (B) and the number of metastatic foci (C). (D) Statistical analysis of IL10 levels in mouse blood. (E) Representative images of HE and IHC staining for SATB2, MIIP, STING, IL10, p52, and CD163 (green arrows) in tumor tissues from the different groups. The bar graphs show the percentages of high/low STING (F), IL-10 (G), and p52 (H) expression in the different groups. (I) Comparison of the number of CD163-positive cells among the different groups. All data are presented as the means ± SDs ( n = 3). ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001; scale bar, 50 μm.

Article Snippet: The primary antibodies used included rabbit anti-MIIP (1:1000, HPA044948; Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-MIIP (1:1000, orb537083; Biorbyt, Cambridge, UK), rabbit anti-STING (1:2000, 19851-1-AP; Proteintech, Wuhan, Hubei, China), mouse anti-TRAF3 (1:1000, 66310-1-Ig; Proteintech, Wuhan, Hubei, China), rabbit anti-p52 (1:1000, 4882S; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-p52 (1:500, 15503-1-AP; Proteintech, Wuhan, Hubei, China), rabbit anti-p-p100 (1:1000, 4810T; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-CD163 (1:1000, 68922; Cell Signaling Technology, Danvers, MA, USA), and mouse anti-β-actin antibodies (1:1000, 3700S; Cell Signaling Technology, Danvers, MA, USA).

Techniques: In Situ, Immunohistochemistry, Expressing, Comparison