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rabbit anti ankrd49 primary antibody  (Proteintech)


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    Proteintech rabbit anti ankrd49 primary antibody
    Fig. 1 <t>ANKRD49</t> is upregulated in NSCLC. (A) The mRNA levels of ANKRD49 in nine fresh NSCLC tissues and corresponding adjacent normal tissues were analyzed by RT-qPCR (N: normal tissues; T: tumorous tissues). (B) The mRNA levels of ANKRD49 were assessed by RT-qPCR in the human bronchial epithelial cell line (HBEC) and seven NSCLC cell lines. Data are expressed as means ± standard deviation. ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001 vs. corresponding adjacent normal lung tissues or HBEC cells
    Rabbit Anti Ankrd49 Primary Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rabbit+anti+ankrd49+primary+antibody/ANKRD49+Antibody/pm37964204-50-0-4
    Average 91 stars, based on 6 article reviews
    rabbit anti ankrd49 primary antibody - by Bioz Stars, 2026-09
    91/100 stars

    Images

    1) Product Images from "ANKRD49 promotes the metastasis of NSCLC via activating JNK-ATF2/c-Jun-MMP-2/9 axis."

    Article Title: ANKRD49 promotes the metastasis of NSCLC via activating JNK-ATF2/c-Jun-MMP-2/9 axis.

    Journal: BMC cancer

    doi: 10.1186/s12885-023-11612-9

    Fig. 1 ANKRD49 is upregulated in NSCLC. (A) The mRNA levels of ANKRD49 in nine fresh NSCLC tissues and corresponding adjacent normal tissues were analyzed by RT-qPCR (N: normal tissues; T: tumorous tissues). (B) The mRNA levels of ANKRD49 were assessed by RT-qPCR in the human bronchial epithelial cell line (HBEC) and seven NSCLC cell lines. Data are expressed as means ± standard deviation. ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001 vs. corresponding adjacent normal lung tissues or HBEC cells
    Figure Legend Snippet: Fig. 1 ANKRD49 is upregulated in NSCLC. (A) The mRNA levels of ANKRD49 in nine fresh NSCLC tissues and corresponding adjacent normal tissues were analyzed by RT-qPCR (N: normal tissues; T: tumorous tissues). (B) The mRNA levels of ANKRD49 were assessed by RT-qPCR in the human bronchial epithelial cell line (HBEC) and seven NSCLC cell lines. Data are expressed as means ± standard deviation. ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001 vs. corresponding adjacent normal lung tissues or HBEC cells

    Techniques Used: Quantitative RT-PCR, Standard Deviation

    Fig. 2 ANKRD49 potentiates migration and invasion of H1299 cells. (A, B) Identification of ANKRD49-OE or ANKRD49-sh H1299 cells was validated by RT- qPCR and Western blot. (C, D) Wound healing assay was used to measure migration of ANKRD49-OE and ANKRD49-sh H1299 cells, representative images were taken at a magnification of 40×, at 0, 24 and 48 h. (E, F) Transwell migration and invasion assays were performed to detect migration and invasion of ANKRD49-OE and ANKRD49-sh H1299 cells. Representative images were taken at 200× magnification. Statistical analysis from five random fields was conducted. All experiments were repeated independently three times. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group, #P < 0.05, ##P < 0.01 vs. LV3 group
    Figure Legend Snippet: Fig. 2 ANKRD49 potentiates migration and invasion of H1299 cells. (A, B) Identification of ANKRD49-OE or ANKRD49-sh H1299 cells was validated by RT- qPCR and Western blot. (C, D) Wound healing assay was used to measure migration of ANKRD49-OE and ANKRD49-sh H1299 cells, representative images were taken at a magnification of 40×, at 0, 24 and 48 h. (E, F) Transwell migration and invasion assays were performed to detect migration and invasion of ANKRD49-OE and ANKRD49-sh H1299 cells. Representative images were taken at 200× magnification. Statistical analysis from five random fields was conducted. All experiments were repeated independently three times. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group, #P < 0.05, ##P < 0.01 vs. LV3 group

    Techniques Used: Migration, Quantitative RT-PCR, Western Blot, Wound Healing Assay, Standard Deviation

    Fig. 3 ANKRD49 upregulates MMP-2/MMP-9 expression in H1299 cells. (A, B) The mRNA and protein levels of MMP-2 and MMP-9 in ANKRD49-OE and ANKRD49-sh H1299 cells were detected using RT-qPCR and Western blot assays. (C, D) The activity of MMP-2 and MMP-9 in ANKRD49-OE and ANKRD49- sh H1299 cells was detected by gelatin zymography. (E, F) A wound healing assay was conducted to assess the effect of MMPs inhibitor (ilomastat) on the migration of ANKRD49-OE H1299 cells; representative images were taken at 40× magnification at 0 and 24 h. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group, #P < 0.05, ##P < 0.01 vs. LV3 group
    Figure Legend Snippet: Fig. 3 ANKRD49 upregulates MMP-2/MMP-9 expression in H1299 cells. (A, B) The mRNA and protein levels of MMP-2 and MMP-9 in ANKRD49-OE and ANKRD49-sh H1299 cells were detected using RT-qPCR and Western blot assays. (C, D) The activity of MMP-2 and MMP-9 in ANKRD49-OE and ANKRD49- sh H1299 cells was detected by gelatin zymography. (E, F) A wound healing assay was conducted to assess the effect of MMPs inhibitor (ilomastat) on the migration of ANKRD49-OE H1299 cells; representative images were taken at 40× magnification at 0 and 24 h. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group, #P < 0.05, ##P < 0.01 vs. LV3 group

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Activity Assay, Zymography, Wound Healing Assay, Migration, Standard Deviation

    Fig. 4 ANKRD49 activates JNK pathway to regulate the expression of MMP-2/MMP-9 in H1299 cells. (A) The MAPK protein levels in ANKRD49-OE and ANKRD49-sh H1299 cells were assessed by Western blot. (B, C) The effects of JNK inhibitor or p38 MAPK inhibitor on the levels of MMP-2/MMP-9 in ANKRD49-OE H1299 cells were tested by Western blot. β-Tubulin served as an internal control. (D, E) A wound healing assay was conducted to assess the effect of JNK inhibitor (SP600125) or p38 inhibitor (SB203580) on the migration of ANKRD49-OE H1299 cells; representative images were taken at 40× magnification at 0 and 24 h. (F) The levels of p-ATF2 and p-c-Jun in ANKRD49-OE and ANKRD49-sh H1299 cells were measured by Western blot. (G, H) The levels of p-ATF2 and p-c-Jun in ANKRD49-OE H1299 cells treated with SP600125, SB203580 or DMSO were detected by Western blot. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group
    Figure Legend Snippet: Fig. 4 ANKRD49 activates JNK pathway to regulate the expression of MMP-2/MMP-9 in H1299 cells. (A) The MAPK protein levels in ANKRD49-OE and ANKRD49-sh H1299 cells were assessed by Western blot. (B, C) The effects of JNK inhibitor or p38 MAPK inhibitor on the levels of MMP-2/MMP-9 in ANKRD49-OE H1299 cells were tested by Western blot. β-Tubulin served as an internal control. (D, E) A wound healing assay was conducted to assess the effect of JNK inhibitor (SP600125) or p38 inhibitor (SB203580) on the migration of ANKRD49-OE H1299 cells; representative images were taken at 40× magnification at 0 and 24 h. (F) The levels of p-ATF2 and p-c-Jun in ANKRD49-OE and ANKRD49-sh H1299 cells were measured by Western blot. (G, H) The levels of p-ATF2 and p-c-Jun in ANKRD49-OE H1299 cells treated with SP600125, SB203580 or DMSO were detected by Western blot. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group

    Techniques Used: Expressing, Western Blot, Control, Wound Healing Assay, Migration, Standard Deviation

    Fig. 5 ANKRD49 activates c-Jun/ATF2 transcription factor to regulate the expression of MMP-2/MMP-9 in H1299 cells. (A) The levels of ATF2 and c-Jun in the nuclear fraction of ANKRD49-OE or LV5 H1299 cells were measured by Western blot. (B) The levels of p-ATF2 and p-c-Jun in the nuclear fraction of ANKRD49-OE or LV5 H1299 cells were measured by Western blot. (C) Co-immunoprecipitation (Co-IP) analysis was performed to assess the interaction between p-ATF2 and p-c-Jun in the nucleus of ANKRD49-OE or vector H1299 cells. (D) Representative immunofluorescence images of ANKRD49 induced nuclear co-location of p-ATF2 and p-c-Jun. Scale bar:100 μm. (E-G) Chromatin immunoprecipitating (CHIP) assay was carried out to analyze the binding of p-ATF2 or p-c-Jun with the promoter of MMP-2 or MMP-9. GAPDH, β-actin, PARP and Histone-H3 served as the internal control for cytosol and nuclear fractions, respectively. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group
    Figure Legend Snippet: Fig. 5 ANKRD49 activates c-Jun/ATF2 transcription factor to regulate the expression of MMP-2/MMP-9 in H1299 cells. (A) The levels of ATF2 and c-Jun in the nuclear fraction of ANKRD49-OE or LV5 H1299 cells were measured by Western blot. (B) The levels of p-ATF2 and p-c-Jun in the nuclear fraction of ANKRD49-OE or LV5 H1299 cells were measured by Western blot. (C) Co-immunoprecipitation (Co-IP) analysis was performed to assess the interaction between p-ATF2 and p-c-Jun in the nucleus of ANKRD49-OE or vector H1299 cells. (D) Representative immunofluorescence images of ANKRD49 induced nuclear co-location of p-ATF2 and p-c-Jun. Scale bar:100 μm. (E-G) Chromatin immunoprecipitating (CHIP) assay was carried out to analyze the binding of p-ATF2 or p-c-Jun with the promoter of MMP-2 or MMP-9. GAPDH, β-actin, PARP and Histone-H3 served as the internal control for cytosol and nuclear fractions, respectively. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group

    Techniques Used: Expressing, Western Blot, Immunoprecipitation, Co-Immunoprecipitation Assay, Plasmid Preparation, Immunofluorescence, Binding Assay, Control, Standard Deviation

    Fig. 6 Knockdown of ANKRD49 inhibits migration, invasion of H1703 cells. (A, B) The stable ANKRD49-sh H1703 cells was validated by RT-qPCR and Western blot. β-actin served as an internal control. (C, D) Wound healing assay was performed to detect migration of ANKRD49-sh H1703 cells, represen tative images were taken at 40× magnification at 0, 24 and 48 h. (E, F) Transwell migration and invasion assays were conducted to evaluate the migration and invasion of ANKRD49-sh H1703 cells. Representative images were taken at 200× magnification. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV3 group
    Figure Legend Snippet: Fig. 6 Knockdown of ANKRD49 inhibits migration, invasion of H1703 cells. (A, B) The stable ANKRD49-sh H1703 cells was validated by RT-qPCR and Western blot. β-actin served as an internal control. (C, D) Wound healing assay was performed to detect migration of ANKRD49-sh H1703 cells, represen tative images were taken at 40× magnification at 0, 24 and 48 h. (E, F) Transwell migration and invasion assays were conducted to evaluate the migration and invasion of ANKRD49-sh H1703 cells. Representative images were taken at 200× magnification. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV3 group

    Techniques Used: Knockdown, Migration, Quantitative RT-PCR, Western Blot, Control, Wound Healing Assay, Standard Deviation

    Fig. 7 Knockdown of ANKRD49 downregulates MMP-2/MMP-9 expression of H1703 cells. (A, B) The mRNA and protein levels of MMP-2 and MMP-9 in ANKRD49-sh H1703 cells were analyzed by RT-qPCR and Western blot. (C, D) The activities of MMP-2 and MMP-9 in ANKRD49-sh H1703 cells were de tected by gelatin zymography. (E) The MAPK proteins in ANKRD49-sh H1703 cells were detected by Western blot. GAPDH served as an internal control. (F) The levels of p-ATF2 and p-c-Jun in ANKRD49-sh H1703 cells were measured by Western blot. (G, H) The levels of JNK, p-JNK, p38, p-p38, MMP-2 and MMP-9 in ANKRD49-sh H1703 cells treated with SP600125, SB203580, Anisomycin or DMSO were measured by Western blot. β-Tubulin served as an inter nal control. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV3 group
    Figure Legend Snippet: Fig. 7 Knockdown of ANKRD49 downregulates MMP-2/MMP-9 expression of H1703 cells. (A, B) The mRNA and protein levels of MMP-2 and MMP-9 in ANKRD49-sh H1703 cells were analyzed by RT-qPCR and Western blot. (C, D) The activities of MMP-2 and MMP-9 in ANKRD49-sh H1703 cells were de tected by gelatin zymography. (E) The MAPK proteins in ANKRD49-sh H1703 cells were detected by Western blot. GAPDH served as an internal control. (F) The levels of p-ATF2 and p-c-Jun in ANKRD49-sh H1703 cells were measured by Western blot. (G, H) The levels of JNK, p-JNK, p38, p-p38, MMP-2 and MMP-9 in ANKRD49-sh H1703 cells treated with SP600125, SB203580, Anisomycin or DMSO were measured by Western blot. β-Tubulin served as an inter nal control. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV3 group

    Techniques Used: Knockdown, Expressing, Quantitative RT-PCR, Western Blot, Zymography, Control, Standard Deviation

    Fig. 8 Overexpression of ANKRD49 promotes the migration and invasion of H1299 cells in nude mice. (A) Representative lung images of mice injected with ANKRD49-OE-H1299 or LV5-H1299 cells. White arrows manifest the metastasis nodules on the lung. (B) Statistical analysis of the number of metas tasis nodules on the lung was illustrated. (C) Representative images of HE staining for lung metastases. Green arrows indicated metastatic H1299 cells. (D) Representative images of IHC staining for NAPSA and NKX2-1. Scale bars represent 100 μm. (E, F) Representative images of IHC staining for ANKRD49, MMP-2, MMP-9, p-JNK, p-ATF2 or p-c-Jun. Scale bars represent 100 μm. (G) Quantitative analysis of IHC staining. **P < 0.01, ***P < 0.001 vs. LV5 group
    Figure Legend Snippet: Fig. 8 Overexpression of ANKRD49 promotes the migration and invasion of H1299 cells in nude mice. (A) Representative lung images of mice injected with ANKRD49-OE-H1299 or LV5-H1299 cells. White arrows manifest the metastasis nodules on the lung. (B) Statistical analysis of the number of metas tasis nodules on the lung was illustrated. (C) Representative images of HE staining for lung metastases. Green arrows indicated metastatic H1299 cells. (D) Representative images of IHC staining for NAPSA and NKX2-1. Scale bars represent 100 μm. (E, F) Representative images of IHC staining for ANKRD49, MMP-2, MMP-9, p-JNK, p-ATF2 or p-c-Jun. Scale bars represent 100 μm. (G) Quantitative analysis of IHC staining. **P < 0.01, ***P < 0.001 vs. LV5 group

    Techniques Used: Over Expression, Migration, Injection, Staining, Immunohistochemistry

    Fig. 9 Knockdown of ANKRD49 declines the migration and invasion of H1703 cells in nude mice. (A) Representative lung images of mice injected with ANKRD49-sh or LV3 H1703 cells. White arrows manifest the metastasis nodules on the lung. (B) Statistical analysis of the number of metastasis nodules on the lung was illustrated. (C) Representative images of HE staining for lung metastases. Green arrows indicated metastatic H1703 cells. (D) Representative images of IHC staining for p63. Scale bars represent 100 μm. (E, F) Representative images of IHC staining for ANKRD49, MMP-2, MMP-9, p-JNK, p-ATF2 or p-c-Jun. Scale bars represent 100 μm. (G) Quantitative analysis of IHC staining. **P < 0.01, ***P < 0.001 vs. LV3 group
    Figure Legend Snippet: Fig. 9 Knockdown of ANKRD49 declines the migration and invasion of H1703 cells in nude mice. (A) Representative lung images of mice injected with ANKRD49-sh or LV3 H1703 cells. White arrows manifest the metastasis nodules on the lung. (B) Statistical analysis of the number of metastasis nodules on the lung was illustrated. (C) Representative images of HE staining for lung metastases. Green arrows indicated metastatic H1703 cells. (D) Representative images of IHC staining for p63. Scale bars represent 100 μm. (E, F) Representative images of IHC staining for ANKRD49, MMP-2, MMP-9, p-JNK, p-ATF2 or p-c-Jun. Scale bars represent 100 μm. (G) Quantitative analysis of IHC staining. **P < 0.01, ***P < 0.001 vs. LV3 group

    Techniques Used: Knockdown, Migration, Injection, Staining, Immunohistochemistry

    Related Articles

    other:

    Article Title: ANKRD49 promotes the metastasis of NSCLC via activating JNK-ATF2/c-Jun-MMP-2/9 axis
    Article Snippet: Rabbit anti-ANKRD49 primary antibody (Proteintech, Cat# 25034-1-AP, RRID:AB_2879860), rabbit anti-MMP-2 polyclonal antibody (Cat# bs-4599R, RRID:AB_11083963), rabbit anti-MMP-9 polyclonal antibody (Cat# bs-0397R, RRID:AB_10853038), rabbit anti-c-Jun polyclonal antibody (Cat# bs-0670R, RRID:AB_10857880), rabbit anti-p-ATF2 polyclonal antibody (Cat# bs-3033R, RRID:AB_10883830) and Alexa Fluor® 488-conjugated goat anti-rabbit IgG (Cat# bs-0295G-A488, RRID:AB_10893781) were obtained from Bioss Biotechnology (Beijing, China).

    Article Title: ANKRD49 promotes the metastasis of NSCLC via activating JNK-ATF2/c-Jun-MMP-2/9 axis.
    Article Snippet: Rabbit anti-ANKRD49 primary antibody (Proteintech, Cat# 25034-1-AP, RRID:AB_2879860), rabbit anti-MMP-2 polyclonal antibody (Cat# bs-4599R, RRID:AB_11083963), rabbit anti-MMP-9 polyclonal antibody (Cat# bs-0397R, RRID:AB_10853038), rabbit anti-c-Jun polyclonal antibody (Cat# bs-0670R, RRID:AB_10857880), rabbit anti-p-ATF2 polyclonal molecular mechanisms of ANKRD49’s function is different from those found in A549 cells.

    Immunohistochemistry:

    Article Title: A Microarray Study on the Expression of ANKRD49 in Lung Squamous Cell Carcinoma and Its Clinicopathologic Significance
    Article Snippet: Lung squamous cell carcinoma (LUSC) is associated with poor clinical outcomes and identifying novel biomarkers that are involved in the progression of LUSC is important for prognosis and targeted treatment.. Herein, ankyrin repeat domain 49 (ANKRD49) protein in LUSC versus paired noncancerous lung tissues was tested and its clinical significance was evaluated through χ test, log-rank test, and Cox proportional hazards model.. The results showed the ANKRD49 protein in LUSC was elevated and correlated with the tumornode-metastasis stage, lymph node metastasis, distal metastasis, and differentiation.

    Staining:

    Article Title: A Microarray Study on the Expression of ANKRD49 in Lung Squamous Cell Carcinoma and Its Clinicopathologic Significance
    Article Snippet: Lung squamous cell carcinoma (LUSC) is associated with poor clinical outcomes and identifying novel biomarkers that are involved in the progression of LUSC is important for prognosis and targeted treatment.. Herein, ankyrin repeat domain 49 (ANKRD49) protein in LUSC versus paired noncancerous lung tissues was tested and its clinical significance was evaluated through χ test, log-rank test, and Cox proportional hazards model.. The results showed the ANKRD49 protein in LUSC was elevated and correlated with the tumornode-metastasis stage, lymph node metastasis, distal metastasis, and differentiation.



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    rabbit anti ankrd49 primary antibody  (Proteintech)
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    Proteintech rabbit anti ankrd49 primary antibody
    Fig. 1 <t>ANKRD49</t> is upregulated in NSCLC. (A) The mRNA levels of ANKRD49 in nine fresh NSCLC tissues and corresponding adjacent normal tissues were analyzed by RT-qPCR (N: normal tissues; T: tumorous tissues). (B) The mRNA levels of ANKRD49 were assessed by RT-qPCR in the human bronchial epithelial cell line (HBEC) and seven NSCLC cell lines. Data are expressed as means ± standard deviation. ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001 vs. corresponding adjacent normal lung tissues or HBEC cells
    Rabbit Anti Ankrd49 Primary Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rabbit+anti+ankrd49+primary+antibody/ANKRD49+Antibody/pm37964204-50-0-4
    Average 91 stars, based on 1 article reviews
    rabbit anti ankrd49 primary antibody - by Bioz Stars, 2026-09
    91/100 stars
    		  Buy from Supplier

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    Fig. 1 ANKRD49 is upregulated in NSCLC. (A) The mRNA levels of ANKRD49 in nine fresh NSCLC tissues and corresponding adjacent normal tissues were analyzed by RT-qPCR (N: normal tissues; T: tumorous tissues). (B) The mRNA levels of ANKRD49 were assessed by RT-qPCR in the human bronchial epithelial cell line (HBEC) and seven NSCLC cell lines. Data are expressed as means ± standard deviation. ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001 vs. corresponding adjacent normal lung tissues or HBEC cells

    Journal: BMC cancer

    Article Title: ANKRD49 promotes the metastasis of NSCLC via activating JNK-ATF2/c-Jun-MMP-2/9 axis.

    doi: 10.1186/s12885-023-11612-9

    Figure Lengend Snippet: Fig. 1 ANKRD49 is upregulated in NSCLC. (A) The mRNA levels of ANKRD49 in nine fresh NSCLC tissues and corresponding adjacent normal tissues were analyzed by RT-qPCR (N: normal tissues; T: tumorous tissues). (B) The mRNA levels of ANKRD49 were assessed by RT-qPCR in the human bronchial epithelial cell line (HBEC) and seven NSCLC cell lines. Data are expressed as means ± standard deviation. ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001 vs. corresponding adjacent normal lung tissues or HBEC cells

    Article Snippet: Rabbit anti-ANKRD49 primary antibody (Proteintech, Cat# 25034-1-AP, RRID:AB_2879860), rabbit anti-MMP-2 polyclonal antibody (Cat# bs-4599R, RRID:AB_11083963), rabbit anti-MMP-9 polyclonal antibody (Cat# bs-0397R, RRID:AB_10853038), rabbit anti-c-Jun polyclonal antibody (Cat# bs-0670R, RRID:AB_10857880), rabbit anti-p-ATF2 polyclonal molecular mechanisms of ANKRD49’s function is different from those found in A549 cells.

    Techniques: Quantitative RT-PCR, Standard Deviation

    Fig. 2 ANKRD49 potentiates migration and invasion of H1299 cells. (A, B) Identification of ANKRD49-OE or ANKRD49-sh H1299 cells was validated by RT- qPCR and Western blot. (C, D) Wound healing assay was used to measure migration of ANKRD49-OE and ANKRD49-sh H1299 cells, representative images were taken at a magnification of 40×, at 0, 24 and 48 h. (E, F) Transwell migration and invasion assays were performed to detect migration and invasion of ANKRD49-OE and ANKRD49-sh H1299 cells. Representative images were taken at 200× magnification. Statistical analysis from five random fields was conducted. All experiments were repeated independently three times. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group, #P < 0.05, ##P < 0.01 vs. LV3 group

    Journal: BMC cancer

    Article Title: ANKRD49 promotes the metastasis of NSCLC via activating JNK-ATF2/c-Jun-MMP-2/9 axis.

    doi: 10.1186/s12885-023-11612-9

    Figure Lengend Snippet: Fig. 2 ANKRD49 potentiates migration and invasion of H1299 cells. (A, B) Identification of ANKRD49-OE or ANKRD49-sh H1299 cells was validated by RT- qPCR and Western blot. (C, D) Wound healing assay was used to measure migration of ANKRD49-OE and ANKRD49-sh H1299 cells, representative images were taken at a magnification of 40×, at 0, 24 and 48 h. (E, F) Transwell migration and invasion assays were performed to detect migration and invasion of ANKRD49-OE and ANKRD49-sh H1299 cells. Representative images were taken at 200× magnification. Statistical analysis from five random fields was conducted. All experiments were repeated independently three times. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group, #P < 0.05, ##P < 0.01 vs. LV3 group

    Article Snippet: Rabbit anti-ANKRD49 primary antibody (Proteintech, Cat# 25034-1-AP, RRID:AB_2879860), rabbit anti-MMP-2 polyclonal antibody (Cat# bs-4599R, RRID:AB_11083963), rabbit anti-MMP-9 polyclonal antibody (Cat# bs-0397R, RRID:AB_10853038), rabbit anti-c-Jun polyclonal antibody (Cat# bs-0670R, RRID:AB_10857880), rabbit anti-p-ATF2 polyclonal molecular mechanisms of ANKRD49’s function is different from those found in A549 cells.

    Techniques: Migration, Quantitative RT-PCR, Western Blot, Wound Healing Assay, Standard Deviation

    Fig. 3 ANKRD49 upregulates MMP-2/MMP-9 expression in H1299 cells. (A, B) The mRNA and protein levels of MMP-2 and MMP-9 in ANKRD49-OE and ANKRD49-sh H1299 cells were detected using RT-qPCR and Western blot assays. (C, D) The activity of MMP-2 and MMP-9 in ANKRD49-OE and ANKRD49- sh H1299 cells was detected by gelatin zymography. (E, F) A wound healing assay was conducted to assess the effect of MMPs inhibitor (ilomastat) on the migration of ANKRD49-OE H1299 cells; representative images were taken at 40× magnification at 0 and 24 h. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group, #P < 0.05, ##P < 0.01 vs. LV3 group

    Journal: BMC cancer

    Article Title: ANKRD49 promotes the metastasis of NSCLC via activating JNK-ATF2/c-Jun-MMP-2/9 axis.

    doi: 10.1186/s12885-023-11612-9

    Figure Lengend Snippet: Fig. 3 ANKRD49 upregulates MMP-2/MMP-9 expression in H1299 cells. (A, B) The mRNA and protein levels of MMP-2 and MMP-9 in ANKRD49-OE and ANKRD49-sh H1299 cells were detected using RT-qPCR and Western blot assays. (C, D) The activity of MMP-2 and MMP-9 in ANKRD49-OE and ANKRD49- sh H1299 cells was detected by gelatin zymography. (E, F) A wound healing assay was conducted to assess the effect of MMPs inhibitor (ilomastat) on the migration of ANKRD49-OE H1299 cells; representative images were taken at 40× magnification at 0 and 24 h. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group, #P < 0.05, ##P < 0.01 vs. LV3 group

    Article Snippet: Rabbit anti-ANKRD49 primary antibody (Proteintech, Cat# 25034-1-AP, RRID:AB_2879860), rabbit anti-MMP-2 polyclonal antibody (Cat# bs-4599R, RRID:AB_11083963), rabbit anti-MMP-9 polyclonal antibody (Cat# bs-0397R, RRID:AB_10853038), rabbit anti-c-Jun polyclonal antibody (Cat# bs-0670R, RRID:AB_10857880), rabbit anti-p-ATF2 polyclonal molecular mechanisms of ANKRD49’s function is different from those found in A549 cells.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Activity Assay, Zymography, Wound Healing Assay, Migration, Standard Deviation

    Fig. 4 ANKRD49 activates JNK pathway to regulate the expression of MMP-2/MMP-9 in H1299 cells. (A) The MAPK protein levels in ANKRD49-OE and ANKRD49-sh H1299 cells were assessed by Western blot. (B, C) The effects of JNK inhibitor or p38 MAPK inhibitor on the levels of MMP-2/MMP-9 in ANKRD49-OE H1299 cells were tested by Western blot. β-Tubulin served as an internal control. (D, E) A wound healing assay was conducted to assess the effect of JNK inhibitor (SP600125) or p38 inhibitor (SB203580) on the migration of ANKRD49-OE H1299 cells; representative images were taken at 40× magnification at 0 and 24 h. (F) The levels of p-ATF2 and p-c-Jun in ANKRD49-OE and ANKRD49-sh H1299 cells were measured by Western blot. (G, H) The levels of p-ATF2 and p-c-Jun in ANKRD49-OE H1299 cells treated with SP600125, SB203580 or DMSO were detected by Western blot. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group

    Journal: BMC cancer

    Article Title: ANKRD49 promotes the metastasis of NSCLC via activating JNK-ATF2/c-Jun-MMP-2/9 axis.

    doi: 10.1186/s12885-023-11612-9

    Figure Lengend Snippet: Fig. 4 ANKRD49 activates JNK pathway to regulate the expression of MMP-2/MMP-9 in H1299 cells. (A) The MAPK protein levels in ANKRD49-OE and ANKRD49-sh H1299 cells were assessed by Western blot. (B, C) The effects of JNK inhibitor or p38 MAPK inhibitor on the levels of MMP-2/MMP-9 in ANKRD49-OE H1299 cells were tested by Western blot. β-Tubulin served as an internal control. (D, E) A wound healing assay was conducted to assess the effect of JNK inhibitor (SP600125) or p38 inhibitor (SB203580) on the migration of ANKRD49-OE H1299 cells; representative images were taken at 40× magnification at 0 and 24 h. (F) The levels of p-ATF2 and p-c-Jun in ANKRD49-OE and ANKRD49-sh H1299 cells were measured by Western blot. (G, H) The levels of p-ATF2 and p-c-Jun in ANKRD49-OE H1299 cells treated with SP600125, SB203580 or DMSO were detected by Western blot. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group

    Article Snippet: Rabbit anti-ANKRD49 primary antibody (Proteintech, Cat# 25034-1-AP, RRID:AB_2879860), rabbit anti-MMP-2 polyclonal antibody (Cat# bs-4599R, RRID:AB_11083963), rabbit anti-MMP-9 polyclonal antibody (Cat# bs-0397R, RRID:AB_10853038), rabbit anti-c-Jun polyclonal antibody (Cat# bs-0670R, RRID:AB_10857880), rabbit anti-p-ATF2 polyclonal molecular mechanisms of ANKRD49’s function is different from those found in A549 cells.

    Techniques: Expressing, Western Blot, Control, Wound Healing Assay, Migration, Standard Deviation

    Fig. 5 ANKRD49 activates c-Jun/ATF2 transcription factor to regulate the expression of MMP-2/MMP-9 in H1299 cells. (A) The levels of ATF2 and c-Jun in the nuclear fraction of ANKRD49-OE or LV5 H1299 cells were measured by Western blot. (B) The levels of p-ATF2 and p-c-Jun in the nuclear fraction of ANKRD49-OE or LV5 H1299 cells were measured by Western blot. (C) Co-immunoprecipitation (Co-IP) analysis was performed to assess the interaction between p-ATF2 and p-c-Jun in the nucleus of ANKRD49-OE or vector H1299 cells. (D) Representative immunofluorescence images of ANKRD49 induced nuclear co-location of p-ATF2 and p-c-Jun. Scale bar:100 μm. (E-G) Chromatin immunoprecipitating (CHIP) assay was carried out to analyze the binding of p-ATF2 or p-c-Jun with the promoter of MMP-2 or MMP-9. GAPDH, β-actin, PARP and Histone-H3 served as the internal control for cytosol and nuclear fractions, respectively. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group

    Journal: BMC cancer

    Article Title: ANKRD49 promotes the metastasis of NSCLC via activating JNK-ATF2/c-Jun-MMP-2/9 axis.

    doi: 10.1186/s12885-023-11612-9

    Figure Lengend Snippet: Fig. 5 ANKRD49 activates c-Jun/ATF2 transcription factor to regulate the expression of MMP-2/MMP-9 in H1299 cells. (A) The levels of ATF2 and c-Jun in the nuclear fraction of ANKRD49-OE or LV5 H1299 cells were measured by Western blot. (B) The levels of p-ATF2 and p-c-Jun in the nuclear fraction of ANKRD49-OE or LV5 H1299 cells were measured by Western blot. (C) Co-immunoprecipitation (Co-IP) analysis was performed to assess the interaction between p-ATF2 and p-c-Jun in the nucleus of ANKRD49-OE or vector H1299 cells. (D) Representative immunofluorescence images of ANKRD49 induced nuclear co-location of p-ATF2 and p-c-Jun. Scale bar:100 μm. (E-G) Chromatin immunoprecipitating (CHIP) assay was carried out to analyze the binding of p-ATF2 or p-c-Jun with the promoter of MMP-2 or MMP-9. GAPDH, β-actin, PARP and Histone-H3 served as the internal control for cytosol and nuclear fractions, respectively. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV5 group

    Article Snippet: Rabbit anti-ANKRD49 primary antibody (Proteintech, Cat# 25034-1-AP, RRID:AB_2879860), rabbit anti-MMP-2 polyclonal antibody (Cat# bs-4599R, RRID:AB_11083963), rabbit anti-MMP-9 polyclonal antibody (Cat# bs-0397R, RRID:AB_10853038), rabbit anti-c-Jun polyclonal antibody (Cat# bs-0670R, RRID:AB_10857880), rabbit anti-p-ATF2 polyclonal molecular mechanisms of ANKRD49’s function is different from those found in A549 cells.

    Techniques: Expressing, Western Blot, Immunoprecipitation, Co-Immunoprecipitation Assay, Plasmid Preparation, Immunofluorescence, Binding Assay, Control, Standard Deviation

    Fig. 6 Knockdown of ANKRD49 inhibits migration, invasion of H1703 cells. (A, B) The stable ANKRD49-sh H1703 cells was validated by RT-qPCR and Western blot. β-actin served as an internal control. (C, D) Wound healing assay was performed to detect migration of ANKRD49-sh H1703 cells, represen tative images were taken at 40× magnification at 0, 24 and 48 h. (E, F) Transwell migration and invasion assays were conducted to evaluate the migration and invasion of ANKRD49-sh H1703 cells. Representative images were taken at 200× magnification. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV3 group

    Journal: BMC cancer

    Article Title: ANKRD49 promotes the metastasis of NSCLC via activating JNK-ATF2/c-Jun-MMP-2/9 axis.

    doi: 10.1186/s12885-023-11612-9

    Figure Lengend Snippet: Fig. 6 Knockdown of ANKRD49 inhibits migration, invasion of H1703 cells. (A, B) The stable ANKRD49-sh H1703 cells was validated by RT-qPCR and Western blot. β-actin served as an internal control. (C, D) Wound healing assay was performed to detect migration of ANKRD49-sh H1703 cells, represen tative images were taken at 40× magnification at 0, 24 and 48 h. (E, F) Transwell migration and invasion assays were conducted to evaluate the migration and invasion of ANKRD49-sh H1703 cells. Representative images were taken at 200× magnification. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV3 group

    Article Snippet: Rabbit anti-ANKRD49 primary antibody (Proteintech, Cat# 25034-1-AP, RRID:AB_2879860), rabbit anti-MMP-2 polyclonal antibody (Cat# bs-4599R, RRID:AB_11083963), rabbit anti-MMP-9 polyclonal antibody (Cat# bs-0397R, RRID:AB_10853038), rabbit anti-c-Jun polyclonal antibody (Cat# bs-0670R, RRID:AB_10857880), rabbit anti-p-ATF2 polyclonal molecular mechanisms of ANKRD49’s function is different from those found in A549 cells.

    Techniques: Knockdown, Migration, Quantitative RT-PCR, Western Blot, Control, Wound Healing Assay, Standard Deviation

    Fig. 7 Knockdown of ANKRD49 downregulates MMP-2/MMP-9 expression of H1703 cells. (A, B) The mRNA and protein levels of MMP-2 and MMP-9 in ANKRD49-sh H1703 cells were analyzed by RT-qPCR and Western blot. (C, D) The activities of MMP-2 and MMP-9 in ANKRD49-sh H1703 cells were de tected by gelatin zymography. (E) The MAPK proteins in ANKRD49-sh H1703 cells were detected by Western blot. GAPDH served as an internal control. (F) The levels of p-ATF2 and p-c-Jun in ANKRD49-sh H1703 cells were measured by Western blot. (G, H) The levels of JNK, p-JNK, p38, p-p38, MMP-2 and MMP-9 in ANKRD49-sh H1703 cells treated with SP600125, SB203580, Anisomycin or DMSO were measured by Western blot. β-Tubulin served as an inter nal control. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV3 group

    Journal: BMC cancer

    Article Title: ANKRD49 promotes the metastasis of NSCLC via activating JNK-ATF2/c-Jun-MMP-2/9 axis.

    doi: 10.1186/s12885-023-11612-9

    Figure Lengend Snippet: Fig. 7 Knockdown of ANKRD49 downregulates MMP-2/MMP-9 expression of H1703 cells. (A, B) The mRNA and protein levels of MMP-2 and MMP-9 in ANKRD49-sh H1703 cells were analyzed by RT-qPCR and Western blot. (C, D) The activities of MMP-2 and MMP-9 in ANKRD49-sh H1703 cells were de tected by gelatin zymography. (E) The MAPK proteins in ANKRD49-sh H1703 cells were detected by Western blot. GAPDH served as an internal control. (F) The levels of p-ATF2 and p-c-Jun in ANKRD49-sh H1703 cells were measured by Western blot. (G, H) The levels of JNK, p-JNK, p38, p-p38, MMP-2 and MMP-9 in ANKRD49-sh H1703 cells treated with SP600125, SB203580, Anisomycin or DMSO were measured by Western blot. β-Tubulin served as an inter nal control. Data are expressed as means ± standard deviation. *P < 0.05, **P < 0.01, ***P < 0.001 vs. LV3 group

    Article Snippet: Rabbit anti-ANKRD49 primary antibody (Proteintech, Cat# 25034-1-AP, RRID:AB_2879860), rabbit anti-MMP-2 polyclonal antibody (Cat# bs-4599R, RRID:AB_11083963), rabbit anti-MMP-9 polyclonal antibody (Cat# bs-0397R, RRID:AB_10853038), rabbit anti-c-Jun polyclonal antibody (Cat# bs-0670R, RRID:AB_10857880), rabbit anti-p-ATF2 polyclonal molecular mechanisms of ANKRD49’s function is different from those found in A549 cells.

    Techniques: Knockdown, Expressing, Quantitative RT-PCR, Western Blot, Zymography, Control, Standard Deviation

    Fig. 8 Overexpression of ANKRD49 promotes the migration and invasion of H1299 cells in nude mice. (A) Representative lung images of mice injected with ANKRD49-OE-H1299 or LV5-H1299 cells. White arrows manifest the metastasis nodules on the lung. (B) Statistical analysis of the number of metas tasis nodules on the lung was illustrated. (C) Representative images of HE staining for lung metastases. Green arrows indicated metastatic H1299 cells. (D) Representative images of IHC staining for NAPSA and NKX2-1. Scale bars represent 100 μm. (E, F) Representative images of IHC staining for ANKRD49, MMP-2, MMP-9, p-JNK, p-ATF2 or p-c-Jun. Scale bars represent 100 μm. (G) Quantitative analysis of IHC staining. **P < 0.01, ***P < 0.001 vs. LV5 group

    Journal: BMC cancer

    Article Title: ANKRD49 promotes the metastasis of NSCLC via activating JNK-ATF2/c-Jun-MMP-2/9 axis.

    doi: 10.1186/s12885-023-11612-9

    Figure Lengend Snippet: Fig. 8 Overexpression of ANKRD49 promotes the migration and invasion of H1299 cells in nude mice. (A) Representative lung images of mice injected with ANKRD49-OE-H1299 or LV5-H1299 cells. White arrows manifest the metastasis nodules on the lung. (B) Statistical analysis of the number of metas tasis nodules on the lung was illustrated. (C) Representative images of HE staining for lung metastases. Green arrows indicated metastatic H1299 cells. (D) Representative images of IHC staining for NAPSA and NKX2-1. Scale bars represent 100 μm. (E, F) Representative images of IHC staining for ANKRD49, MMP-2, MMP-9, p-JNK, p-ATF2 or p-c-Jun. Scale bars represent 100 μm. (G) Quantitative analysis of IHC staining. **P < 0.01, ***P < 0.001 vs. LV5 group

    Article Snippet: Rabbit anti-ANKRD49 primary antibody (Proteintech, Cat# 25034-1-AP, RRID:AB_2879860), rabbit anti-MMP-2 polyclonal antibody (Cat# bs-4599R, RRID:AB_11083963), rabbit anti-MMP-9 polyclonal antibody (Cat# bs-0397R, RRID:AB_10853038), rabbit anti-c-Jun polyclonal antibody (Cat# bs-0670R, RRID:AB_10857880), rabbit anti-p-ATF2 polyclonal molecular mechanisms of ANKRD49’s function is different from those found in A549 cells.

    Techniques: Over Expression, Migration, Injection, Staining, Immunohistochemistry

    Fig. 9 Knockdown of ANKRD49 declines the migration and invasion of H1703 cells in nude mice. (A) Representative lung images of mice injected with ANKRD49-sh or LV3 H1703 cells. White arrows manifest the metastasis nodules on the lung. (B) Statistical analysis of the number of metastasis nodules on the lung was illustrated. (C) Representative images of HE staining for lung metastases. Green arrows indicated metastatic H1703 cells. (D) Representative images of IHC staining for p63. Scale bars represent 100 μm. (E, F) Representative images of IHC staining for ANKRD49, MMP-2, MMP-9, p-JNK, p-ATF2 or p-c-Jun. Scale bars represent 100 μm. (G) Quantitative analysis of IHC staining. **P < 0.01, ***P < 0.001 vs. LV3 group

    Journal: BMC cancer

    Article Title: ANKRD49 promotes the metastasis of NSCLC via activating JNK-ATF2/c-Jun-MMP-2/9 axis.

    doi: 10.1186/s12885-023-11612-9

    Figure Lengend Snippet: Fig. 9 Knockdown of ANKRD49 declines the migration and invasion of H1703 cells in nude mice. (A) Representative lung images of mice injected with ANKRD49-sh or LV3 H1703 cells. White arrows manifest the metastasis nodules on the lung. (B) Statistical analysis of the number of metastasis nodules on the lung was illustrated. (C) Representative images of HE staining for lung metastases. Green arrows indicated metastatic H1703 cells. (D) Representative images of IHC staining for p63. Scale bars represent 100 μm. (E, F) Representative images of IHC staining for ANKRD49, MMP-2, MMP-9, p-JNK, p-ATF2 or p-c-Jun. Scale bars represent 100 μm. (G) Quantitative analysis of IHC staining. **P < 0.01, ***P < 0.001 vs. LV3 group

    Article Snippet: Rabbit anti-ANKRD49 primary antibody (Proteintech, Cat# 25034-1-AP, RRID:AB_2879860), rabbit anti-MMP-2 polyclonal antibody (Cat# bs-4599R, RRID:AB_11083963), rabbit anti-MMP-9 polyclonal antibody (Cat# bs-0397R, RRID:AB_10853038), rabbit anti-c-Jun polyclonal antibody (Cat# bs-0670R, RRID:AB_10857880), rabbit anti-p-ATF2 polyclonal molecular mechanisms of ANKRD49’s function is different from those found in A549 cells.

    Techniques: Knockdown, Migration, Injection, Staining, Immunohistochemistry