Journal: Cell Communication and Signaling : CCS

Article Title: HPV11 targeting KDM4A regulates the polarization of macrophage M 1 and promotes the development of nasal inverted papilloma

doi: 10.1186/s12964-024-01971-6

Figure Lengend Snippet: The primers for qRT-PCR

Article Snippet: Primary antibodies used were: anti- KDM4A (1:5000, Proteintech, Wuhan, China), anti-HPV11E6 (1:1000, MyBioSource, San Diego, USA), anti-HPV11E7 (1:1000, Abcam, Shanghai, China), anti-CD206 (1:1000, Proteintech, Wuhan, China), anti-CD86 (1:2000, Proteintech, Wuhan, China), anti-NF-κB P65 (1:5000, Proteintech, Wuhan, China), anti-Phospho-NF-κB P65 (1:5000, Proteintech, Wuhan, China), anti-Phospho-IκB-α (1:10000, Proteintech, Wuhan, China) and anti-GAPDH (1:20000, Proteintech, Wuhan, China).

Techniques:

Journal: Cell Communication and Signaling : CCS

Article Title: HPV11 targeting KDM4A regulates the polarization of macrophage M 1 and promotes the development of nasal inverted papilloma

doi: 10.1186/s12964-024-01971-6

Figure Lengend Snippet: A Pupil map of integration sites of HPV ( +) viral genes in the human chromosome genome in NIP: each short band marked on the human chromosome in the figure indicates an integration event, and the high-frequency integration site KDM4A for HPV infection was identified. B-D The mRNA expression of HPV11E6, HPV11E7, and KDM4A in the cells of each group was verified by qRT-PCR. E , F The protein expression of HPV11E6, HPV11E7, and KDM4A in cells of each group was verified by Western blot. G - I Image J was used to analyze the gray values of each strip. Data are presented as mean ± SD; *** P < 0.001, ** P < 0.002, * P < 0.033, ns means no statistical significance ( P > 0.12)

Article Snippet: Primary antibodies used were: anti- KDM4A (1:5000, Proteintech, Wuhan, China), anti-HPV11E6 (1:1000, MyBioSource, San Diego, USA), anti-HPV11E7 (1:1000, Abcam, Shanghai, China), anti-CD206 (1:1000, Proteintech, Wuhan, China), anti-CD86 (1:2000, Proteintech, Wuhan, China), anti-NF-κB P65 (1:5000, Proteintech, Wuhan, China), anti-Phospho-NF-κB P65 (1:5000, Proteintech, Wuhan, China), anti-Phospho-IκB-α (1:10000, Proteintech, Wuhan, China) and anti-GAPDH (1:20000, Proteintech, Wuhan, China).

Techniques: Infection, Expressing, Quantitative RT-PCR, Western Blot, Stripping Membranes

Journal: Cell Communication and Signaling : CCS

Article Title: HPV11 targeting KDM4A regulates the polarization of macrophage M 1 and promotes the development of nasal inverted papilloma

doi: 10.1186/s12964-024-01971-6

Figure Lengend Snippet: Overexpression of HPV11E6/E7 can promote cell proliferation and migration in vitro, and KDM4A gene knockout can inhibit cell proliferation and migration caused by HPV11E6/E7 overexpression. A Cell proliferation was measured by the CCK-8 method. B , C Colony formation assay was used to detect cell proliferation. D , E EdU fluorescent labeling was used to detect cell proliferation. F , G Transwell assay was used to detect cell migration. H , I Wound healing test to detect cell migration. Data are presented as mean ± SD; *** P < 0.001, ** P < 0.002, * P < 0.033

Article Snippet: Primary antibodies used were: anti- KDM4A (1:5000, Proteintech, Wuhan, China), anti-HPV11E6 (1:1000, MyBioSource, San Diego, USA), anti-HPV11E7 (1:1000, Abcam, Shanghai, China), anti-CD206 (1:1000, Proteintech, Wuhan, China), anti-CD86 (1:2000, Proteintech, Wuhan, China), anti-NF-κB P65 (1:5000, Proteintech, Wuhan, China), anti-Phospho-NF-κB P65 (1:5000, Proteintech, Wuhan, China), anti-Phospho-IκB-α (1:10000, Proteintech, Wuhan, China) and anti-GAPDH (1:20000, Proteintech, Wuhan, China).

Techniques: Over Expression, Migration, In Vitro, Gene Knockout, CCK-8 Assay, Colony Assay, Labeling, Transwell Assay

Journal: Cell Communication and Signaling : CCS

Article Title: HPV11 targeting KDM4A regulates the polarization of macrophage M 1 and promotes the development of nasal inverted papilloma

doi: 10.1186/s12964-024-01971-6

Figure Lengend Snippet: Overexpression of HPV11E6/E7 can promote M 1 polarization of macrophages under co-culture conditions, and knockout of KDM4A can reduce the degree of M 1 polarization of macrophages. A - C Typical markers of M 1 macrophages (CD206, IL-6, IL-1β) were detected by qRT-PCR. D - F Typical markers of M 2 macrophages (CD206, IL-10, Arg-1) were detected by qRT-PCR. G - K Western blot was used to detect the expression levels of CD86 and CD206 proteins and the expressions of NF-κB P65, Phospho-NF-κB P65, and Phospho-IκB proteins in the M 1 macrophage polarization pathway NF-kB. The internal parameter of CD86, CD206 and Phospho-IκB is GAPDH, and the internal parameter of Phospho-NF-κB P65 is NF-κB P65. *** P < 0.001, ** P < 0.002, * P < 0.033, ns means no statistical significance( P > 0.12)

Article Snippet: Primary antibodies used were: anti- KDM4A (1:5000, Proteintech, Wuhan, China), anti-HPV11E6 (1:1000, MyBioSource, San Diego, USA), anti-HPV11E7 (1:1000, Abcam, Shanghai, China), anti-CD206 (1:1000, Proteintech, Wuhan, China), anti-CD86 (1:2000, Proteintech, Wuhan, China), anti-NF-κB P65 (1:5000, Proteintech, Wuhan, China), anti-Phospho-NF-κB P65 (1:5000, Proteintech, Wuhan, China), anti-Phospho-IκB-α (1:10000, Proteintech, Wuhan, China) and anti-GAPDH (1:20000, Proteintech, Wuhan, China).

Techniques: Over Expression, Co-Culture Assay, Knock-Out, Quantitative RT-PCR, Western Blot, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: HPV11 targeting KDM4A regulates the polarization of macrophage M 1 and promotes the development of nasal inverted papilloma

doi: 10.1186/s12964-024-01971-6

Figure Lengend Snippet: In vivo experiments have shown that overexpression of HPV11E6/E7 can promote subcutaneous tumor formation in nude mice, and KDM4A knockout can inhibit the proliferation of subcutaneous tumor tissue. Overexpression of HPV11E6/E7 can promote local recruitment of more macrophages and induce M 1 -type polarization of macrophages in subcutaneous tumors of nude mice. Knockout of KDM4A gene can reduce the number of macrophages recruited and the degree of M 1 -type polarization of macrophages. A , B The volume of subcutaneous mass tissue in each group in the nude mouse model. C - E The expression level of HPV11E6, HPV11E7, and KDM4A in subcutaneous mass tissues of mice in each group was verified by qRT-PCR. F H&E staining of subcutaneous tumor tissue. G Tissue immunofluorescence F4/80 + CD86 double staining. H Tissue immunofluorescence F4/80 + CD206 double staining. I-K Semi-quantitative analysis of the average fluorescence intensity of F4/80, CD86, and CD206 in each group. Data are presented as mean ± SD; *** P < 0.001, ** P < 0.002, ns means no statistical significance( P > 0.12)

Article Snippet: Primary antibodies used were: anti- KDM4A (1:5000, Proteintech, Wuhan, China), anti-HPV11E6 (1:1000, MyBioSource, San Diego, USA), anti-HPV11E7 (1:1000, Abcam, Shanghai, China), anti-CD206 (1:1000, Proteintech, Wuhan, China), anti-CD86 (1:2000, Proteintech, Wuhan, China), anti-NF-κB P65 (1:5000, Proteintech, Wuhan, China), anti-Phospho-NF-κB P65 (1:5000, Proteintech, Wuhan, China), anti-Phospho-IκB-α (1:10000, Proteintech, Wuhan, China) and anti-GAPDH (1:20000, Proteintech, Wuhan, China).

Techniques: In Vivo, Over Expression, Knock-Out, Expressing, Quantitative RT-PCR, Staining, Immunofluorescence, Double Staining, Fluorescence