rabbit anti basic fgf  (Cell Signaling Technology Inc)

Journal: Biology Open

Article Title: miR-302 regulates pancreatic progenitor pool and pancreatic size

doi: 10.1242/bio.062353

Figure Lengend Snippet: mir-302 regulates pancreatic size rather than cellular composition. (A,B) Immunofluorescence (IF) staining of (A) INSULIN and (B) GLUCAGON in WT and KO pancreas at E17.5. The liver, pancreas, spleen, and intestine were dissected together from the embryos, and embedded and sectioned together coronally. The pancreas was outlined in white dashed line. Scale bars: 500 μm. (C) Quantification of pancreatic size outlined in dashed line in A and B. (D) IF staining of BrdU (green) and ECAD (yellow) after 30-min pulse before dissection at E17.5. Scale bar: 50 μm. (E) Quantification of BrdU+ cells in the pancreas in D. (F) Zoomed in images of INSULIN and GLUCAGON staining showing their localization in islets. Scale bars: 50 μm. (G) Quantification of INSULIN+ and GLUCAGON+ cells in the pancreas in A and B. (H) IF staining of MUCIN1 (green) and E-CADHERIN (yellow) showing their localization in the acini at E17.5. Scale bar: 50 μm. (I) Quantification of MUCIN1+ in the pancreas in H. P -value in (C,E,G,I) by Student’s t -test. Each data point represented an embryo quantified with at least 12 coronal sections spaced equally across the pancreas.

Article Snippet: Primary antibodies used in this study and their dilutions were: GFP (ab13970, Abcam, 1:500), Pdx1 (F6A11, DSHB, 1:400), insulin (sc-8033, Santa Cruz Biotechnology, 1:500), glucagon (sc-514562, Santa Cruz Biotechnology, 1:500), E-cadherin (3196, Cell Signaling Technology, 1:200), mucin1 (MA5-11202, Invitrogen, 1:500), BrdU (ab6326, Abcam, 1:500), phospho-histone H3 (9701, Cell Signaling Technology, 1:500).

Techniques: Immunofluorescence, Staining, Dissection

Journal: Cell Communication and Signaling : CCS

Article Title: Senescent cells promote breast cancer cells motility by secreting GM-CSF and bFGF that activate the JNK signaling pathway

doi: 10.1186/s12964-024-01861-x

Figure Lengend Snippet: Senescent cells secrete bFGF and GM-CSF that promote the motility of breast cancer cells ( A ) Relative mRNA levels (mean ± SD, n = 3) of GM-CSF, bFGF and CXCL1 in young (PD31) and senescent (PD62) BJ cells as determined by quantitative real time PCR analysis. ( B ) Relative protein levels (mean ± SD, n = 3) of GM-CSF, bFGF and CXCL1 in the conditioned medium from young (PD31) and senescent (PD62) BJ cells as determined by ELISA. ( C - D ) Representative crystal violet-stained images of transwell migration of MDA-MB-231, MDA-MB-468 and MCF-7 cells co-cultured with medium containing DMSO, 10 ng/ml of bFGF, 20 ng/ml of GM-CSF or both for 12 h ( C ), and quantification of number of migrated cells per field (mean ± SD, n = 3) ( D ). At least 5 randomly chosen 20X fields were counted for each of the triplicates. ( E - F ) Representative images of MDA-MB-231, MDA-MB-468 and MCF-7 cells immediately (0 h), 24 h (24 h, for MDA-MB-231 and MDA-MB-468) or 48 h (48 h, for MCF-7) after a scratch wound was made and co-cultured with medium containing DMSO, 10 ng/ml of bFGF, 20 ng/ml of GM-CSF or both ( E ), and quantification of the distance of the wound edges by ImageJ at 24 h (for MDA-MB-231 and MDA-MB-468) or 48 h (for MCF-7) (mean ± SD, n = 3) ( F ). ( A - B , D , F ) * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. BJ-PD31 in unpaired, 2-sample t-tests ( A - B ) or between indicated groups in One-way ANOVA corrected for multiple comparisons using Tukey’s multiple comparison correction adjustment ( D , F )

Article Snippet: Neutralizing antibodies against bFGF (Cell Signaling, 98658) and GM-CSF (Cell Signaling, 56712) were added to the medium of BJ cells at the final concentration of 1 ng/mL and 5 ng/mL, respectively.

Techniques: Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Staining, Migration, Cell Culture, Comparison

Journal: Cell Communication and Signaling : CCS

Article Title: Senescent cells promote breast cancer cells motility by secreting GM-CSF and bFGF that activate the JNK signaling pathway

doi: 10.1186/s12964-024-01861-x

Figure Lengend Snippet: Senescent cells promote the migration of breast cancer cells via secreted bFGF and GM-CSF. ( A - B ) Representative crystal violet-stained images of transwell migration of MDA-MB-231, MDA-MB-468 and MCF-7 cells co-cultured with conditioned medium from young (PD29) or senescent (PD61) BJ cells, which were left untreated (None) or incubated with neutralizing antibodies against β-actin (1 ng/ml), bFGF (1 ng/ml), GM-CSF (5 ng/ml) or both bFGF and GM-CSF for 12 h ( A ), and quantification of number of migrated cells per field (mean ± SD, n = 3) ( B ). At least 5 randomly chosen 20X fields were counted for each of the triplicates ( C - D ) Representative crystal violet-stained images of transwell migration of MDA-MB-231, MDA-MB-468 and MCF-7 cells co-cultured with conditioned medium from young (PD29) or senescent (PD61) BJ cells transduced with shRNA control (SC) or shRNAs for bFGF or GM-CSF for 20 h ( C ), and quantification of number of migrated cells per field (mean ± SD, n = 3) ( D ). At least 5 randomly chosen 20X fields were counted for each of the triplicates. ( B , D ) ns, not significant; * p < 0.05; ** p < 0.01; and *** p < 0.001 between indicated groups in unpaired, 2-sample t tests (dotted lines) or One-way ANOVA corrected for multiple comparisons using Tukey’s multiple comparison correction adjustment (solid lines)

Article Snippet: Neutralizing antibodies against bFGF (Cell Signaling, 98658) and GM-CSF (Cell Signaling, 56712) were added to the medium of BJ cells at the final concentration of 1 ng/mL and 5 ng/mL, respectively.

Techniques: Migration, Staining, Cell Culture, Incubation, Transduction, shRNA, Control, Comparison

Journal: Cell Communication and Signaling : CCS

Article Title: Senescent cells promote breast cancer cells motility by secreting GM-CSF and bFGF that activate the JNK signaling pathway

doi: 10.1186/s12964-024-01861-x

Figure Lengend Snippet: Senescent cells induce JNK activation via secreted GMCSF and bFGF ( A - B ) Western blotting analysis of the phosphorylation/activation status of Stat3, AKT, ERK and JNK in MDA-MB-231, MDA-MB-468 and MCF-7 breast cancer cell lines co-cultured with medium (None) or young (PD31) or senescent (PD56) BJ cells. ( C - D ) Western blotting analysis of phosphorylated/activated JNK in MDA-MB-231 and MDA-MB-468 breast cancer cell lines co-cultured with medium (None) or young (PD31) or senescent (PD59) BJ cells, which were incubated with neutralizing antibodies against IgG, GM-CSF (αG, 5 ng/ml), bFGF (αF, 1 ng/ml), or both bFGF and GM-CSF (αG + αF). ( E - F ) Western blotting analysis of phosphorylated/activated JNK in MDA-MB-231 and MDA-MB-468 breast cancer cell lines treated with GM-CSF, bFGF or both. ( B , D , F ) Quantification of the Western blot results presented in A, C and E, respectively, showing expression levels of indicated proteins relative to None ( B ), None + IgG ( D ) or Ctrl ( F ). Densitometric quantification of bands in the Western blots was performed by ImageJ. The signals were normalized to that of actin

Article Snippet: Neutralizing antibodies against bFGF (Cell Signaling, 98658) and GM-CSF (Cell Signaling, 56712) were added to the medium of BJ cells at the final concentration of 1 ng/mL and 5 ng/mL, respectively.

Techniques: Activation Assay, Western Blot, Phospho-proteomics, Cell Culture, Incubation, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: Senescent cells promote breast cancer cells motility by secreting GM-CSF and bFGF that activate the JNK signaling pathway

doi: 10.1186/s12964-024-01861-x

Figure Lengend Snippet: JNK mediates the stimulation of breast cancer migration by senescent cell-secreted GM-CSF and bFGF ( A ) Western blotting analysis showing knockdown specificity and efficiency of JNK1 and JNK2 shRNAs in MDA-MB-231 and MDA-MB-468 cells. ( B - C ) Representative crystal violet-stained images of transwell migration of MDA-MB-231 and MDA-MB-468 cells co-cultured with young (PD31) or senescent (PD59) BJ cells transduced with shRNA control (SC) or shRNAs for JNK1 or JNK2 for 12 h ( C ), and quantification of number of migrated cells per field (mean ± SD, n = 3) ( D ). At least 5 randomly chosen 20X fields were counted for each of the triplicates. ( D - E ) Representative crystal violet-stained images of transwell migration of MDA-MB-231 cells transduced with shRNAs for JNK1 or JNK2 or shRNA control (SC) and treated with vehicle control, 20 ng/ml of GM-CSF, 10 ng/ml of bFGF or both for 12 h ( D ), and quantification of number per field (mean ± SD, n = 3) of migrated MDA-MB-231 and MDA-MB-468 cells transduced with shRNAs for JNK1 or JNK2 or shRNA control (SC) and treated with vehicle control, 20 ng/ml of GM-CSF, 10 ng/ml of bFGF or both for 13 h ( E ). At least 5 randomly chosen 20X fields were counted for each of the triplicates. ( C , E ) ** p < 0.01 and *** p < 0.001 between indicated groups in unpaired, 2-sample t tests (dotted lines) or One-way ANOVA corrected for multiple comparisons using Tukey’s multiple comparison correction adjustment (solid lines)

Article Snippet: Neutralizing antibodies against bFGF (Cell Signaling, 98658) and GM-CSF (Cell Signaling, 56712) were added to the medium of BJ cells at the final concentration of 1 ng/mL and 5 ng/mL, respectively.

Techniques: Migration, Western Blot, Knockdown, Staining, Cell Culture, Transduction, shRNA, Control, Comparison

Journal: Cell Communication and Signaling : CCS

Article Title: Senescent cells promote breast cancer cells motility by secreting GM-CSF and bFGF that activate the JNK signaling pathway

doi: 10.1186/s12964-024-01861-x

Figure Lengend Snippet: Senescent cells secrete bFGF and GM-CSF that promote the motility of breast cancer cells ( A ) Relative mRNA levels (mean ± SD, n = 3) of GM-CSF, bFGF and CXCL1 in young (PD31) and senescent (PD62) BJ cells as determined by quantitative real time PCR analysis. ( B ) Relative protein levels (mean ± SD, n = 3) of GM-CSF, bFGF and CXCL1 in the conditioned medium from young (PD31) and senescent (PD62) BJ cells as determined by ELISA. ( C - D ) Representative crystal violet-stained images of transwell migration of MDA-MB-231, MDA-MB-468 and MCF-7 cells co-cultured with medium containing DMSO, 10 ng/ml of bFGF, 20 ng/ml of GM-CSF or both for 12 h ( C ), and quantification of number of migrated cells per field (mean ± SD, n = 3) ( D ). At least 5 randomly chosen 20X fields were counted for each of the triplicates. ( E - F ) Representative images of MDA-MB-231, MDA-MB-468 and MCF-7 cells immediately (0 h), 24 h (24 h, for MDA-MB-231 and MDA-MB-468) or 48 h (48 h, for MCF-7) after a scratch wound was made and co-cultured with medium containing DMSO, 10 ng/ml of bFGF, 20 ng/ml of GM-CSF or both ( E ), and quantification of the distance of the wound edges by ImageJ at 24 h (for MDA-MB-231 and MDA-MB-468) or 48 h (for MCF-7) (mean ± SD, n = 3) ( F ). ( A - B , D , F ) * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. BJ-PD31 in unpaired, 2-sample t-tests ( A - B ) or between indicated groups in One-way ANOVA corrected for multiple comparisons using Tukey’s multiple comparison correction adjustment ( D , F )

Article Snippet: The following primary antibodies and concentration were used: β-actin (Santa Cruz, sc-47778 1:10000), JNK1 (Abcam, ab199380 1:1000), JNK2 (Cell Signaling, 9258 S 1:500), E-cadherin (BD Biosciences, 610181 1:1000), Vimentin (Cell Signaling, 5741 S 1:1000), p-AKT(Ser473) (Cell Signaling, 9271 S 1:1000), p-JNK (Thr183/Tyr185) (Cell Signaling, 9251 S 1:1000), JNK (Cell Signaling, 9252 1:1000), bFGF (Cell Signaling, 98658 S 1:1000), GM-CSF (Abcam, ab300495 1:1000), p-Stat3 (Ser727) (Santa Cruz, sc-8001-R, 1:1000), p-Stat3 (Tyr705) (B7, Santa Cruz, sc-8059, 1:1000), p-ERK1/2 (Thr202/Tyr204) (BIOSS, bs-3016R, 1:1000), ERK1/2 (C9, Santa Cruz, sc-514302, 1:1000).

Techniques: Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Staining, Migration, Cell Culture, Comparison

Journal: Cell Communication and Signaling : CCS

Article Title: Senescent cells promote breast cancer cells motility by secreting GM-CSF and bFGF that activate the JNK signaling pathway

doi: 10.1186/s12964-024-01861-x

Figure Lengend Snippet: Senescent cells promote the migration of breast cancer cells via secreted bFGF and GM-CSF. ( A - B ) Representative crystal violet-stained images of transwell migration of MDA-MB-231, MDA-MB-468 and MCF-7 cells co-cultured with conditioned medium from young (PD29) or senescent (PD61) BJ cells, which were left untreated (None) or incubated with neutralizing antibodies against β-actin (1 ng/ml), bFGF (1 ng/ml), GM-CSF (5 ng/ml) or both bFGF and GM-CSF for 12 h ( A ), and quantification of number of migrated cells per field (mean ± SD, n = 3) ( B ). At least 5 randomly chosen 20X fields were counted for each of the triplicates ( C - D ) Representative crystal violet-stained images of transwell migration of MDA-MB-231, MDA-MB-468 and MCF-7 cells co-cultured with conditioned medium from young (PD29) or senescent (PD61) BJ cells transduced with shRNA control (SC) or shRNAs for bFGF or GM-CSF for 20 h ( C ), and quantification of number of migrated cells per field (mean ± SD, n = 3) ( D ). At least 5 randomly chosen 20X fields were counted for each of the triplicates. ( B , D ) ns, not significant; * p < 0.05; ** p < 0.01; and *** p < 0.001 between indicated groups in unpaired, 2-sample t tests (dotted lines) or One-way ANOVA corrected for multiple comparisons using Tukey’s multiple comparison correction adjustment (solid lines)

Article Snippet: The following primary antibodies and concentration were used: β-actin (Santa Cruz, sc-47778 1:10000), JNK1 (Abcam, ab199380 1:1000), JNK2 (Cell Signaling, 9258 S 1:500), E-cadherin (BD Biosciences, 610181 1:1000), Vimentin (Cell Signaling, 5741 S 1:1000), p-AKT(Ser473) (Cell Signaling, 9271 S 1:1000), p-JNK (Thr183/Tyr185) (Cell Signaling, 9251 S 1:1000), JNK (Cell Signaling, 9252 1:1000), bFGF (Cell Signaling, 98658 S 1:1000), GM-CSF (Abcam, ab300495 1:1000), p-Stat3 (Ser727) (Santa Cruz, sc-8001-R, 1:1000), p-Stat3 (Tyr705) (B7, Santa Cruz, sc-8059, 1:1000), p-ERK1/2 (Thr202/Tyr204) (BIOSS, bs-3016R, 1:1000), ERK1/2 (C9, Santa Cruz, sc-514302, 1:1000).

Techniques: Migration, Staining, Cell Culture, Incubation, Transduction, shRNA, Control, Comparison

Journal: Cell Communication and Signaling : CCS

Article Title: Senescent cells promote breast cancer cells motility by secreting GM-CSF and bFGF that activate the JNK signaling pathway

doi: 10.1186/s12964-024-01861-x

Figure Lengend Snippet: Senescent cells induce JNK activation via secreted GMCSF and bFGF ( A - B ) Western blotting analysis of the phosphorylation/activation status of Stat3, AKT, ERK and JNK in MDA-MB-231, MDA-MB-468 and MCF-7 breast cancer cell lines co-cultured with medium (None) or young (PD31) or senescent (PD56) BJ cells. ( C - D ) Western blotting analysis of phosphorylated/activated JNK in MDA-MB-231 and MDA-MB-468 breast cancer cell lines co-cultured with medium (None) or young (PD31) or senescent (PD59) BJ cells, which were incubated with neutralizing antibodies against IgG, GM-CSF (αG, 5 ng/ml), bFGF (αF, 1 ng/ml), or both bFGF and GM-CSF (αG + αF). ( E - F ) Western blotting analysis of phosphorylated/activated JNK in MDA-MB-231 and MDA-MB-468 breast cancer cell lines treated with GM-CSF, bFGF or both. ( B , D , F ) Quantification of the Western blot results presented in A, C and E, respectively, showing expression levels of indicated proteins relative to None ( B ), None + IgG ( D ) or Ctrl ( F ). Densitometric quantification of bands in the Western blots was performed by ImageJ. The signals were normalized to that of actin

Article Snippet: The following primary antibodies and concentration were used: β-actin (Santa Cruz, sc-47778 1:10000), JNK1 (Abcam, ab199380 1:1000), JNK2 (Cell Signaling, 9258 S 1:500), E-cadherin (BD Biosciences, 610181 1:1000), Vimentin (Cell Signaling, 5741 S 1:1000), p-AKT(Ser473) (Cell Signaling, 9271 S 1:1000), p-JNK (Thr183/Tyr185) (Cell Signaling, 9251 S 1:1000), JNK (Cell Signaling, 9252 1:1000), bFGF (Cell Signaling, 98658 S 1:1000), GM-CSF (Abcam, ab300495 1:1000), p-Stat3 (Ser727) (Santa Cruz, sc-8001-R, 1:1000), p-Stat3 (Tyr705) (B7, Santa Cruz, sc-8059, 1:1000), p-ERK1/2 (Thr202/Tyr204) (BIOSS, bs-3016R, 1:1000), ERK1/2 (C9, Santa Cruz, sc-514302, 1:1000).

Techniques: Activation Assay, Western Blot, Phospho-proteomics, Cell Culture, Incubation, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: Senescent cells promote breast cancer cells motility by secreting GM-CSF and bFGF that activate the JNK signaling pathway

doi: 10.1186/s12964-024-01861-x

Figure Lengend Snippet: JNK mediates the stimulation of breast cancer migration by senescent cell-secreted GM-CSF and bFGF ( A ) Western blotting analysis showing knockdown specificity and efficiency of JNK1 and JNK2 shRNAs in MDA-MB-231 and MDA-MB-468 cells. ( B - C ) Representative crystal violet-stained images of transwell migration of MDA-MB-231 and MDA-MB-468 cells co-cultured with young (PD31) or senescent (PD59) BJ cells transduced with shRNA control (SC) or shRNAs for JNK1 or JNK2 for 12 h ( C ), and quantification of number of migrated cells per field (mean ± SD, n = 3) ( D ). At least 5 randomly chosen 20X fields were counted for each of the triplicates. ( D - E ) Representative crystal violet-stained images of transwell migration of MDA-MB-231 cells transduced with shRNAs for JNK1 or JNK2 or shRNA control (SC) and treated with vehicle control, 20 ng/ml of GM-CSF, 10 ng/ml of bFGF or both for 12 h ( D ), and quantification of number per field (mean ± SD, n = 3) of migrated MDA-MB-231 and MDA-MB-468 cells transduced with shRNAs for JNK1 or JNK2 or shRNA control (SC) and treated with vehicle control, 20 ng/ml of GM-CSF, 10 ng/ml of bFGF or both for 13 h ( E ). At least 5 randomly chosen 20X fields were counted for each of the triplicates. ( C , E ) ** p < 0.01 and *** p < 0.001 between indicated groups in unpaired, 2-sample t tests (dotted lines) or One-way ANOVA corrected for multiple comparisons using Tukey’s multiple comparison correction adjustment (solid lines)

Article Snippet: The following primary antibodies and concentration were used: β-actin (Santa Cruz, sc-47778 1:10000), JNK1 (Abcam, ab199380 1:1000), JNK2 (Cell Signaling, 9258 S 1:500), E-cadherin (BD Biosciences, 610181 1:1000), Vimentin (Cell Signaling, 5741 S 1:1000), p-AKT(Ser473) (Cell Signaling, 9271 S 1:1000), p-JNK (Thr183/Tyr185) (Cell Signaling, 9251 S 1:1000), JNK (Cell Signaling, 9252 1:1000), bFGF (Cell Signaling, 98658 S 1:1000), GM-CSF (Abcam, ab300495 1:1000), p-Stat3 (Ser727) (Santa Cruz, sc-8001-R, 1:1000), p-Stat3 (Tyr705) (B7, Santa Cruz, sc-8059, 1:1000), p-ERK1/2 (Thr202/Tyr204) (BIOSS, bs-3016R, 1:1000), ERK1/2 (C9, Santa Cruz, sc-514302, 1:1000).

Techniques: Migration, Western Blot, Knockdown, Staining, Cell Culture, Transduction, shRNA, Control, Comparison

Journal: Frontiers in Pharmacology

Article Title: Scaling up production of recombinant human basic fibroblast growth factor in an Escherichia coli BL21(DE3) plysS strain and evaluation of its pro-wound healing efficacy

doi: 10.3389/fphar.2023.1279516

Figure Lengend Snippet: Recombinant plasmid construction and expression of the hbFGF protein. (A) Schematic diagram of the mpET3c-hbFGF recombinant plasmid construction. (B) Kanamycin resistance detection of the mpET3c-hbFGF recombinant plasmid. 1, E. coli BL21 (DE3) plysS-pET3c strain. 2, E. coli BL21 (DE3) plysS-mpET3c/hbFGF engineered strain. (C) Restriction enzyme analysis of the mpET3c-hbFGF recombinant plasmid. Lanes 1 and 4, DNA molecular weight marker. Lane 2, plasmid before digestion. Lane 3, plasmid after digestion. (D) SDS-PAGE (left) and Western blot (right) analyses of the expressed hbFGF protein in E. coli BL21(DE3) plysS. Lanes 1 and 4, non-induced. Lanes 3 and 5, induced by IPTG for 4 h. Lane M, molecular weight marker. Black arrows indicate hbFGF.

Article Snippet: The polyclonal rabbit anti-human bFGF antibody (Cat.No: 3196S) was acquired from Cell Signaling Technology (United States), while the secondary antibody (goat anti-rabbit lgG/HRP antibody, Cat.No: HS101-01) was procured from Transgene Biotech (Beijing, China).

Techniques: Recombinant, Plasmid Preparation, Expressing, Molecular Weight, Marker, SDS Page, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Scaling up production of recombinant human basic fibroblast growth factor in an Escherichia coli BL21(DE3) plysS strain and evaluation of its pro-wound healing efficacy

doi: 10.3389/fphar.2023.1279516

Figure Lengend Snippet: Tested cultural conditions for hbFGF production on a flask scale.

Article Snippet: The polyclonal rabbit anti-human bFGF antibody (Cat.No: 3196S) was acquired from Cell Signaling Technology (United States), while the secondary antibody (goat anti-rabbit lgG/HRP antibody, Cat.No: HS101-01) was procured from Transgene Biotech (Beijing, China).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Scaling up production of recombinant human basic fibroblast growth factor in an Escherichia coli BL21(DE3) plysS strain and evaluation of its pro-wound healing efficacy

doi: 10.3389/fphar.2023.1279516

Figure Lengend Snippet: Optimizing of fermentation parameters of hbFGF E. coli strain in 30 mL of LB medium. (A) The 12-h growth curve of the hbFGF E. coli strain in 30 mL of LB medium at 37°C and 200 rpm. (B–E) The 12-h growth curve of hbFGF E. coli strain under different conditions, including (B) glucose concentrations in the range of 0.5–20 g/L, (C) pH in the range of 6.6–7.4, (D) temperatures of 32°C, 34°C, 36°C, and 38°C, and (E) medium volume (30, 50, 75, and 100 mL). (F) The expression level of hbFGF (f1) and bacterial density (f2) after induction at different OD 600 values with 0.8 mM IPTG for 1–6 h. (G) The 10-h growth curve of the hbFGF E. coli strain under different inoculations. All experiments were performed in a 250-mL shake flask. Asterisks indicate significant difference (* p < 0.05, ** p < 0.01, *** p < 0.001, n = 3).

Article Snippet: The polyclonal rabbit anti-human bFGF antibody (Cat.No: 3196S) was acquired from Cell Signaling Technology (United States), while the secondary antibody (goat anti-rabbit lgG/HRP antibody, Cat.No: HS101-01) was procured from Transgene Biotech (Beijing, China).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Scaling up production of recombinant human basic fibroblast growth factor in an Escherichia coli BL21(DE3) plysS strain and evaluation of its pro-wound healing efficacy

doi: 10.3389/fphar.2023.1279516

Figure Lengend Snippet: Three-dimensional response surface for the bacterial density and hbFGF expression level. (A) Effects on the bacterial density of (a1) temperature and pH; (a2) temperature and IPTG concentration; (a3) temperature and NH 4 Cl concentration; (a4) temperature and induction time; (a5) pH and IPTG concentration; (a6) pH and NH 4 Cl concentration; (a7) pH and induction time; (a8) IPTG concentration and NH 4 Cl concentration; (a9) IPTG concentration and induction time; and (a10) NH 4 Cl concentration and induction time. (B) Effect on the hbFGF expression level of (b1) temperature and pH; (b2) temperature and IPTG concentration; (b3) temperature and NH 4 Cl concentration; and (b4) temperature and induction time.

Article Snippet: The polyclonal rabbit anti-human bFGF antibody (Cat.No: 3196S) was acquired from Cell Signaling Technology (United States), while the secondary antibody (goat anti-rabbit lgG/HRP antibody, Cat.No: HS101-01) was procured from Transgene Biotech (Beijing, China).

Techniques: Expressing, Concentration Assay

Journal: Frontiers in Pharmacology

Article Title: Scaling up production of recombinant human basic fibroblast growth factor in an Escherichia coli BL21(DE3) plysS strain and evaluation of its pro-wound healing efficacy

doi: 10.3389/fphar.2023.1279516

Figure Lengend Snippet: The optimal induction conditions for the hbFGF fermentation.

Article Snippet: The polyclonal rabbit anti-human bFGF antibody (Cat.No: 3196S) was acquired from Cell Signaling Technology (United States), while the secondary antibody (goat anti-rabbit lgG/HRP antibody, Cat.No: HS101-01) was procured from Transgene Biotech (Beijing, China).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Scaling up production of recombinant human basic fibroblast growth factor in an Escherichia coli BL21(DE3) plysS strain and evaluation of its pro-wound healing efficacy

doi: 10.3389/fphar.2023.1279516

Figure Lengend Snippet: Summary of scale-up fermentation data for hbFGF (Mean ± SD, n ≥ 3).

Article Snippet: The polyclonal rabbit anti-human bFGF antibody (Cat.No: 3196S) was acquired from Cell Signaling Technology (United States), while the secondary antibody (goat anti-rabbit lgG/HRP antibody, Cat.No: HS101-01) was procured from Transgene Biotech (Beijing, China).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Scaling up production of recombinant human basic fibroblast growth factor in an Escherichia coli BL21(DE3) plysS strain and evaluation of its pro-wound healing efficacy

doi: 10.3389/fphar.2023.1279516

Figure Lengend Snippet: Process of fermentation of E. coli BL21 (DE3) plysS-mpET3c/hbFGF. (A) Growth curve of hbFGF engineered strain in a 200-L fermentation. (B) SDS-PAGE analysis of the hbFGF expression at the 500-L scale. #1 Lane, non-induced. Lane M, molecular weight marker. (C) Variations in the parameters for hbFGF production at the 500-L scale, including wet cell concentration (OD 600 ), temperature, dissolved oxygen (DO) levels, rotation speed, and air ventilation rate. (D) The relative curves of the specific growth rate, pH, glucose addition, and nitrogen source addition with time in a 500-L fermentation. Black arrows indicate hbFGF.

Article Snippet: The polyclonal rabbit anti-human bFGF antibody (Cat.No: 3196S) was acquired from Cell Signaling Technology (United States), while the secondary antibody (goat anti-rabbit lgG/HRP antibody, Cat.No: HS101-01) was procured from Transgene Biotech (Beijing, China).

Techniques: SDS Page, Expressing, Molecular Weight, Marker, Concentration Assay

Journal: Frontiers in Pharmacology

Article Title: Scaling up production of recombinant human basic fibroblast growth factor in an Escherichia coli BL21(DE3) plysS strain and evaluation of its pro-wound healing efficacy

doi: 10.3389/fphar.2023.1279516

Figure Lengend Snippet: Summary of the purification process for hbFGF (Mean ± SD, n = 4).

Article Snippet: The polyclonal rabbit anti-human bFGF antibody (Cat.No: 3196S) was acquired from Cell Signaling Technology (United States), while the secondary antibody (goat anti-rabbit lgG/HRP antibody, Cat.No: HS101-01) was procured from Transgene Biotech (Beijing, China).

Techniques: Purification, Bacteria, Lysis

Journal: Frontiers in Pharmacology

Article Title: Scaling up production of recombinant human basic fibroblast growth factor in an Escherichia coli BL21(DE3) plysS strain and evaluation of its pro-wound healing efficacy

doi: 10.3389/fphar.2023.1279516

Figure Lengend Snippet: Identification and analysis of hbFGF in the purification process. (A) SDS-PAGE analysis of hbFGF after ion exchange and affinity chromatography. Lane 1, supernatant. Lane 2, the flow-through sample from CM-Sepharose. Lane 3, 0.36 M NaCl-eluted sample from CM-Sepharose. Lane 4, 2.0 M NaCl-regenerated sample from CM-Sepharose. Lane 5, flow-through fraction from heparin affinity Sepharose. Lane 6, 2.0 M NaCl-eluted sample from heparin affinity Sepharose. Lane 7, 2.0 M NaCl-regenerated sample from SP-Sepharose. Lane 8, purified hbFGF eluted with 0.5 M NaCl from SP-Sepharose. Lane M, molecular weight marker. (B) RP-HPLC and (C) SEC-HPLC analysis of purified hbFGF. (D) The biological activity of hbFGF on NIH-3T3 cells. (E) Analysis of Isoelectric point, (F) Mass spectrum, (G) molecular peptide mapping coverage, and (H) CD spectrum analysis of purified hbFGF. Black arrows indicate hbFGF.

Article Snippet: The polyclonal rabbit anti-human bFGF antibody (Cat.No: 3196S) was acquired from Cell Signaling Technology (United States), while the secondary antibody (goat anti-rabbit lgG/HRP antibody, Cat.No: HS101-01) was procured from Transgene Biotech (Beijing, China).

Techniques: Purification, SDS Page, Affinity Chromatography, Molecular Weight, Marker, Activity Assay

Journal: Frontiers in Pharmacology

Article Title: Scaling up production of recombinant human basic fibroblast growth factor in an Escherichia coli BL21(DE3) plysS strain and evaluation of its pro-wound healing efficacy

doi: 10.3389/fphar.2023.1279516

Figure Lengend Snippet: Comparison of hbFGF expressed for various hosts.

Article Snippet: The polyclonal rabbit anti-human bFGF antibody (Cat.No: 3196S) was acquired from Cell Signaling Technology (United States), while the secondary antibody (goat anti-rabbit lgG/HRP antibody, Cat.No: HS101-01) was procured from Transgene Biotech (Beijing, China).

Techniques: Comparison

Journal: Frontiers in Pharmacology

Article Title: Scaling up production of recombinant human basic fibroblast growth factor in an Escherichia coli BL21(DE3) plysS strain and evaluation of its pro-wound healing efficacy

doi: 10.3389/fphar.2023.1279516

Figure Lengend Snippet: The healing effect of hbFGF on a deep second-degree scald wound model in rats. (A) Photos of the skin wound and wound healing rates of STZ-induced SD rats with deep second-degree scald wounds. (B) Results of Masson (scale bar, 200 μm) and IHC staining (scale bar, 50 μm) and the pathological score of a deep second-degree scald wound model in rats. (C) The expression level of FGF-1, TGF-β1, and hydroxyproline in a deep second-degree scald wound model in rats. Compared with the control group, * p < 0.05, ** p < 0.01, *** p < 0.001, n = 7.

Article Snippet: The polyclonal rabbit anti-human bFGF antibody (Cat.No: 3196S) was acquired from Cell Signaling Technology (United States), while the secondary antibody (goat anti-rabbit lgG/HRP antibody, Cat.No: HS101-01) was procured from Transgene Biotech (Beijing, China).

Techniques: Immunohistochemistry, Expressing, Control

Journal: iScience

Article Title: Exosomes derived from platelet-rich plasma promote diabetic wound healing via the JAK2/STAT3 pathway

doi: 10.1016/j.isci.2023.108236

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-Basic FGF , Cell Signaling Technology , Cat#98658; RRID: AB_398188.

Techniques: Recombinant, cDNA Synthesis, Software