human bmp7 elisa kit  (Proteintech)

Journal: Cancer Science

Article Title: p53 Deficiency in Colon Cancer Cells Promotes Tumor Progression Through the Modulation of Meflin in Fibroblasts

doi: 10.1111/cas.70026

Figure Lengend Snippet: TP53 inactivation in cancer cells suppresses Meflin expression in fibroblasts. (A and B) Relative mRNA (A) and protein (B) levels of TP53 in HCT116sh control and HCT116sh p53 cells; n = 3; Student's t ‐test, * p < 0.05. (C) Viability of HCT116sh control or HCT116sh p53 cells cocultured with or without CCD‐18Co cells for 72 h, as assessed using the WST assay. Only cancer cells were assessed, without including CCD‐18Co cells; n = 3; two‐way ANOVA, * p < 0.05. (D) Tumor volume in BALB/c nude mice subcutaneously injected with HCT116sh control or HCT116sh p53 cells, with or without CCD‐18Co cells; n = 10, two‐way repeated measures ANOVA; * p < 0.05, compared to the volume in mice injected with HCT116sh control cells together with CCD‐18Co cells. (E) Relative mRNA levels of Meflin, TGF‐β1, VEGFA, and BMP7 in CCD‐18Co cells cocultured with or without HCT116sh control or HCT116sh p53 cells; n = 3; one‐way ANOVA, * p < 0.05. (F) Western blot analysis of Meflin expression in CCD‐18Co cells cocultured with or without HCT116sh control or HCT116sh p53 cells.

Article Snippet: TGF‐β levels in cell culture supernatants were measured using a human TGF‐β and BMP7 Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Control, WST Assay, Injection, Western Blot

Journal: Cancer Science

Article Title: p53 Deficiency in Colon Cancer Cells Promotes Tumor Progression Through the Modulation of Meflin in Fibroblasts

doi: 10.1111/cas.70026

Figure Lengend Snippet: Meflin expression in fibroblasts is regulated by TGF‐β and vitamin D. (A) Relative mRNA levels of TGF‐β1 , VEGFA , and Meflin in CCD‐18Co cells treated with or without recombinant TGF‐β (10 ng/mL); n = 3; Student's t ‐test, * p < 0.05. (B) Western blotting for Meflin in CCD‐18Co cells treated with recombinant TGF‐β. (C and D) TGF‐β and BMP7 levels in the supernatant of HCT116 sh control and HCT116 sh p53 cells cultured alone (C) or together with CCD‐18Co cells (D) were analyzed using ELISA. Equal amounts of HCT116 and CCD‐18Co cells were cultured for 48 h in six‐well plates; n = 4; Student's t ‐test; * p < 0.05. (E and F) Relative mRNA (E) and protein (F) levels of Meflin in CCD‐18Co cells treated with or without recombinant calcitriol (100 nM); n = 3; Student's t ‐test, * p < 0.05.

Article Snippet: TGF‐β levels in cell culture supernatants were measured using a human TGF‐β and BMP7 Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Recombinant, Western Blot, Control, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Induced retinal pigment epithelial cells with anti-epithelial-to-mesenchymal transition ability delay retinal degeneration.

doi: 10.1016/j.isci.2022.105050

Figure Lengend Snippet: Figure 2. BMP7 and FOXF2 are critical regulators of EMT in iRPE cells (A) The higher level of BMP7 secreted by iRPE cells compared with De-iPSC-RPE cells was determined by ELISA. Data are mean G SD, unpaired two-sided t-tests, n = 4. (B and C) The reduced expression level of FOXF2 in iRPE compared with that in De-iPSC-RPE cells was determined by (B) immunostaining and (C) western blotting. Scale bar = 50 mm.

Article Snippet: BMP7 was quantified by human BMP7 ELISA kit (R&D Systems, Minneapolis, MN). iPSC-RPE cells, De-iPSC-RPE cells, and iRPE cells were cultured in 24-transwell plate for one month, the supernatants from upper and lower chambers of 24-transwell plate were collected.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Immunostaining, Western Blot

Journal: iScience

Article Title: Induced retinal pigment epithelial cells with anti-epithelial-to-mesenchymal transition ability delay retinal degeneration.

doi: 10.1016/j.isci.2022.105050

Figure Lengend Snippet: Figure 6. Four TFs transcriptionally regulate bmp7, foxf2, lin7a, pard6b, and ppm1a in a direct or indirect manner (A) ELISA analysis demonstrated that BMP7 levels was reduced in 4TFsnr2e1-RPE, 4TFsmitf-a-RPE, 4TFsc-myc-RPE, and 4TFscrx-RPE cells compared with that in iRPE cells. Data are mean G SD, one-way ANOVA and post hoc Bonferroni test, n = 4. (B and C) The levels of FOXF2, LIN7A, PARD6B, and PPM1A were determined by (B) western blotting and (C) quantitative analysis. Data are mean G SD, one- way ANOVA and post hoc Bonferroni test, n = 3. (D) Generation of FLAG-MITF-A-iRPE, FLAG-CRX-iRPE, FLAG-NR2E1-iRPE, and FLAG-C-MYC-iRPE cells. Scale bar = 50 mm. (E–I) Enriched peaks of CRX binding to the target genes and fold enrichment of CRX immunoprecipitation compared with IgG control, as determined by qRT-PCR. Data are mean G SD, unpaired two-sided t-tests, n = 3. (J–M) Enriched peaks of MITF-A and NR2E1 binding to the target gene lin7a and fold enrichment of MITF-A and NR2E1 immunoprecipitation compared with IgG control, as determined by qRT-PCR. Data are mean G SD, unpaired two-sided t-tests, n = 3. (N) Schematic model for the regulation of EMT and MET processes by the four TFs. CRX, MITF-A, NR2E1, and C-MYC directly or indirectly regulated the expression of bmp7, lin7a, pard6b, ppm1a, and foxf2 to inhibit EMT and promote MET.

Article Snippet: BMP7 was quantified by human BMP7 ELISA kit (R&D Systems, Minneapolis, MN). iPSC-RPE cells, De-iPSC-RPE cells, and iRPE cells were cultured in 24-transwell plate for one month, the supernatants from upper and lower chambers of 24-transwell plate were collected.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Binding Assay, Immunoprecipitation, Control, Quantitative RT-PCR, Expressing

Journal: iScience

Article Title: Induced retinal pigment epithelial cells with anti-epithelial-to-mesenchymal transition ability delay retinal degeneration.

doi: 10.1016/j.isci.2022.105050

Figure Lengend Snippet: Figure 2. BMP7 and FOXF2 are critical regulators of EMT in iRPE cells (A) The higher level of BMP7 secreted by iRPE cells compared with De-iPSC-RPE cells was determined by ELISA. Data are mean G SD, unpaired two-sided t-tests, n = 4. (B and C) The reduced expression level of FOXF2 in iRPE compared with that in De-iPSC-RPE cells was determined by (B) immunostaining and (C) western blotting. Scale bar = 50 mm.

Article Snippet: BMP7 was quantified by human BMP7 ELISA kit (R&D Systems, Minneapolis, MN). iPSC-RPE cells, De-iPSC-RPE cells, and iRPE cells were cultured in 24-transwell plate for one month, the supernatants from upper and lower chambers of 24-transwell plate were collected.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Immunostaining, Western Blot

Journal: iScience

Article Title: Induced retinal pigment epithelial cells with anti-epithelial-to-mesenchymal transition ability delay retinal degeneration.

doi: 10.1016/j.isci.2022.105050

Figure Lengend Snippet: Figure 6. Four TFs transcriptionally regulate bmp7, foxf2, lin7a, pard6b, and ppm1a in a direct or indirect manner (A) ELISA analysis demonstrated that BMP7 levels was reduced in 4TFsnr2e1-RPE, 4TFsmitf-a-RPE, 4TFsc-myc-RPE, and 4TFscrx-RPE cells compared with that in iRPE cells. Data are mean G SD, one-way ANOVA and post hoc Bonferroni test, n = 4. (B and C) The levels of FOXF2, LIN7A, PARD6B, and PPM1A were determined by (B) western blotting and (C) quantitative analysis. Data are mean G SD, one- way ANOVA and post hoc Bonferroni test, n = 3. (D) Generation of FLAG-MITF-A-iRPE, FLAG-CRX-iRPE, FLAG-NR2E1-iRPE, and FLAG-C-MYC-iRPE cells. Scale bar = 50 mm. (E–I) Enriched peaks of CRX binding to the target genes and fold enrichment of CRX immunoprecipitation compared with IgG control, as determined by qRT-PCR. Data are mean G SD, unpaired two-sided t-tests, n = 3. (J–M) Enriched peaks of MITF-A and NR2E1 binding to the target gene lin7a and fold enrichment of MITF-A and NR2E1 immunoprecipitation compared with IgG control, as determined by qRT-PCR. Data are mean G SD, unpaired two-sided t-tests, n = 3. (N) Schematic model for the regulation of EMT and MET processes by the four TFs. CRX, MITF-A, NR2E1, and C-MYC directly or indirectly regulated the expression of bmp7, lin7a, pard6b, ppm1a, and foxf2 to inhibit EMT and promote MET.

Article Snippet: BMP7 was quantified by human BMP7 ELISA kit (R&D Systems, Minneapolis, MN). iPSC-RPE cells, De-iPSC-RPE cells, and iRPE cells were cultured in 24-transwell plate for one month, the supernatants from upper and lower chambers of 24-transwell plate were collected.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Binding Assay, Immunoprecipitation, Control, Quantitative RT-PCR, Expressing