anti fabp5 rabbit polyclonal antibody  (Proteintech)

Journal: Cell reports

Article Title: Keratinocyte FABP5-VCP complex mediates recruitment of neutrophils in psoriasis

doi: 10.1016/j.celrep.2023.113449

Figure Lengend Snippet: (A) FABP5 levels in skin samples from healthy individuals and patients with psoriasis. Data were analyzed by publicly accessing database GSE14905 from the Gene Expression Omnibus (GEO). The database of GSE14905 contained 21 healthy individuals and 33 psoriasis patients. (B) FABP5 levels in skin samples from healthy individuals and patients with psoriasis. Data were analyzed by publicly accessing GEO database GSE13355, which contained 64 healthy individuals and 58 psoriasis patients. (C) Alexa Fluor 488-labeled FABP5 expression profile in human skin tissue collected from psoriasis patients (lower panels) and healthy individuals (upper panels). H&E staining (left panels) showed the structure of skin tissue from psoriasis patients and healthy individuals. (D) Fabp5 mRNA levels in skin tissues from imiquimod (IMQ)-induced psoriasis mice and normal mice (n = 8/group). (E) H&E staining (left panels) and immunohistochemistry staining (right panels) showed the structure of skin tissue and Fabp5 levels from IMQ-induced psoriasis mice and control mice, respectively. Data are shown as mean ± SD in (A, B, and D) (**p ≤ 0.01, ****p ≤ 0.0001 as compared with healthy or control groups, unpaired Student’s t test). See also .

Article Snippet: Rabbit polyclonal anti-human FABP5 , Invitrogen , Cat#PA5-80612, RRID:AB_2787908.

Techniques: Expressing, Labeling, Staining, Immunohistochemistry, Control

Journal: Cell reports

Article Title: Keratinocyte FABP5-VCP complex mediates recruitment of neutrophils in psoriasis

doi: 10.1016/j.celrep.2023.113449

Figure Lengend Snippet: (A) Representative images of skin from Fabp5 global knockout and WT mice after treatment with IMQ for 7 days. On day 7 and WT controls. (B) The score of erythema, induration, and desquamation on skin were evaluated from the first day on both Fabp5 global knockout mice and WT controls. The cumulative score was calculated in a sum of the score of erythema, induration, and desquamation at the indicated time point (n = 5/group). (C) Ear thickness was measured daily for both strains, and its change was calculated by subtracting the first day value (n = 5/group). (D and E) H&E staining for ear (D) and skin (E) from both Fabp5 global knockout mice and WT controls. (F–H) Neutrophil infiltration and ratio were analyzed from ear (F), dermis (G), and PBMC (H) using flow cytometry at the endpoint of treatment for both Fabp5 global knockout mice and WT controls. In each figure, left panel: representative flow plot with gate on neutrophils (Ly6G + ) from Fabp5 global knockout mice and WT controls. The parent population is CD45 + Zombie − ; right panel: histogram represents neutrophil ratio in different skin tissues (n = 5/group). Data are shown as mean ± SD in (B, C, and F–H) (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001 as compared with healthy or control groups, unpaired Student’s t test). See also .

Article Snippet: Rabbit polyclonal anti-human FABP5 , Invitrogen , Cat#PA5-80612, RRID:AB_2787908.

Techniques: Knock-Out, Staining, Flow Cytometry, Control

Journal: Cell reports

Article Title: Keratinocyte FABP5-VCP complex mediates recruitment of neutrophils in psoriasis

doi: 10.1016/j.celrep.2023.113449

Figure Lengend Snippet: (A) Representative images of the skin from Fabp5 f/f Krt6A-Cre + and Fabp5 f/f Krt6A-Cre − mice after treatment with IMQ for 7 days. (B) The cumulative score (a sum score of erythema, induration, and desquamation on skin) was calculated from the first day on both Fabp5 f/f Krt6A-Cre + and Fabp5 f/f Krt6A-Cre − mice (n = 10/group). (C) Ear thickness was measured daily from Fabp5 f/f Krt6A-Cre + and Fabp5 f/f Krt6A-Cre − , and its change was calculated by subtracting the first day value (n = 10/group). (D) Neutrophil infiltration and its ratio were analyzed from the ear using flow cytometry at the endpoint of treatment for both Fabp5 f/f Krt6A-Cre + and Fabp5 f/f Krt6A-Cre − mice. In each figure, left panel: representative flow plot with gate on neutrophils (Ly6G + ) from Fabp5 global knockout mice and WT controls. The parent population is CD45 + Zombie − ; right panel: histogram represents neutrophil ratio in different tissues (n = 6/group). (E and F) Neutrophils infiltration and ratio were analyzed from the ear (D), dermis (E), and PBMC (F) using flow cytometry at the endpoint of treatment for both Fabp5 f/f Krt6A-Cre + and Fabp5 f/f Krt6A-Cre − mice. In each figure, left is the flow plot with gate on neutrophils (Ly6G + ) in Fabp5 global knockout and WT mice. The parent population is CD45 + Zombie − ; right is the histogram showing neutrophil ratio in individual tissues (n = 8/group). (G) Images were taken from skin with IMQ-induced psoriasis on day 7 from Fabp5 f/f LysM-Cre + and Fabp5 f/f LysM-Cre − . (H) Ear thickness was measured daily from Fabp5 f/f LysM-Cre + and Fabp5 f/f LysM-Cre – , and its change was calculated by subtracting the first day value (n = 6/group). (I) The cumulative score (a sum score of erythema, induration, and desquamation on skin) was calculated from the first day on both Fabp5 f/f LysM-Cre + and Fabp5 f/f LysM-Cre – mice (n = 6/group). (J–L) Neutrophils infiltration and its ratio was analyzed from the ear (J), dermis (K), and PBMC (L) using flow cytometry at the endpoint of treatment for both Fabp5 f/f LysM-Cre + and Fabp5 f/f LysM-Cre – mice. In each panel, flow cytometric plot (left) showed neutrophils (Ly6G + ) population in Fabp5 global knockout and WT mice; histogram (right) represented neutrophil ratio in CD45 + immune cells in individual tissues (n = 7/group). Data are shown as mean ± SD in (D–F, K, and L) (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001; ns, not significant compared with control groups, unpaired Student’s t test). See also and .

Article Snippet: Rabbit polyclonal anti-human FABP5 , Invitrogen , Cat#PA5-80612, RRID:AB_2787908.

Techniques: Flow Cytometry, Knock-Out, Control

Journal: Cell reports

Article Title: Keratinocyte FABP5-VCP complex mediates recruitment of neutrophils in psoriasis

doi: 10.1016/j.celrep.2023.113449

Figure Lengend Snippet: (A–D) Real-time PCR showed downregulation of Fabp5 (A), IL-36γ (B), Cxcl1 (C), and Cxcl2 (D) with IMQ-treated skin tissue for 24 h in Fabp5 global knockout mice compared with WT controls (n = 3/group). (E–J) Real-time PCR showed the levels of FABP5 (E), CXCL1 (F), CXCL2 (G), CXCL8 (H), IL-36γ (I), and KLK6 (J) were downregulated in FABP5-deficient HaCaT cells transfected with 40 nM siRNA compared with siNC controls with 10 ng/mL TNF-α, 50 μg/mL IMQ, and DMSO as control treatment for 6 h (n = 5/group). (K) Immunohistochemistry staining on skin tissues from two representative psoriasis patients showed FABP5 expression (left) and neutrophil elastase expression (right). Data are shown as mean ± SD in (A–J) (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001; ns, not significant compared with control groups, unpaired Student’s t test). See also .

Article Snippet: Rabbit polyclonal anti-human FABP5 , Invitrogen , Cat#PA5-80612, RRID:AB_2787908.

Techniques: Real-time Polymerase Chain Reaction, Knock-Out, Transfection, Control, Immunohistochemistry, Staining, Expressing

Journal: Cell reports

Article Title: Keratinocyte FABP5-VCP complex mediates recruitment of neutrophils in psoriasis

doi: 10.1016/j.celrep.2023.113449

Figure Lengend Snippet: (A) The workflow on immunoprecipitation with anti-FABP5 antibody in HaCaT cells. (B and C) Silver staining (B) and Coomassie blue staining (C) on SDS-PAGE gel for immunoprecipitated complexes. Blue arrow shows the band for FABP5, and orange arrow shows the band for VCP. (D and E) Validation of FABP5-VCP interaction using immunoprecipitation and western blot. HaCaT lysates were immunoprecipitated (IP) with anti-FABP5 antibody (D) and anti-VCP antibody (E), separately. Immunoblots were performed with anti-VCP antibody and anti-FABP5 antibody as indicated in the figure. (F and G) Images of HaCaT cells show co-localization of FABP5 and VCP using confocal analysis (F). The straight line indicates the region of interest (ROI) utilized to measure the fluorescence intensity in both VCP and FABP5 (G). FABP5 (green), VCP (red), Hochest33342 (blue). Scale bars, 50 μm. (H) Detection of NF-κB signaling activation using western blot. Transient silencing FABP5 with RNAi for 24 h in HaCaT cells and controls were treated with 10 ng/mL TNF-α for the indicated time points. FABP5, VCP, p-IκBα(S32), and p-NFκB(S536) were detected. β-Actin was used as internal control. (I–K) The levels of p-IκBα(S32), p-NFκB p65 (S536), and FABP5 were quantified using ImageJ from three independent western blots experiments (*p < 0.05). Data are shown as mean ± SD in (I–K) (*p ≤ 0.05 compared with the siNC control group, unpaired Student’s t test). See also .

Article Snippet: Rabbit polyclonal anti-human FABP5 , Invitrogen , Cat#PA5-80612, RRID:AB_2787908.

Techniques: Immunoprecipitation, Silver Staining, Staining, SDS Page, Western Blot, Fluorescence, Activation Assay, Control

Journal: Cell reports

Article Title: Keratinocyte FABP5-VCP complex mediates recruitment of neutrophils in psoriasis

doi: 10.1016/j.celrep.2023.113449

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-human FABP5 , Invitrogen , Cat#PA5-80612, RRID:AB_2787908.

Techniques: Control, Recombinant, Activation Assay, Staining, Cream, Reverse Transcription, Polymer, Plasmid Preparation, Blocking Assay, Real-time Polymerase Chain Reaction, SYBR Green Assay, Microarray, Software

Journal: PLOS ONE

Article Title: Fatty acid binding protein 5 regulates docetaxel sensitivity in taxane-resistant prostate cancer cells

doi: 10.1371/journal.pone.0292483

Figure Lengend Snippet: ( A ) qPCR and ( B ) western blotting demonstrates FABP5 expression in DU145 and DU145-TXR cells and confirms successful FABP5 KD in the corresponding cell-lines (n = 3). ( C ) Quantification of western blots presented as FABP5 / β-Actin (n = 3). ( D ) Cytotoxicity of docetaxel in taxane-resistant DU145-TXR and taxane-sensitive DU145 cells and in cells bearing an FABP5 KD. The IC 50 values for docetaxel cytotoxicity were 6.1 nM, 5.3 nM, >100 nM, and 4.4 nM in DU145, DU145 KD, DU145-TXR, and DU145-TXR KD cells, respectively. **, p < 0.01; ***, p < 0.001 (n = 4). Data are reported as % viability of the respective vehicle controls.

Article Snippet: The following antibodies were used: mouse anti-β-actin (Cell Signaling, #3700), rabbit anti-FABP5 (BioVendor, #RD181060100), and anti-ABCB1 (Cell Signaling, #12683).

Techniques: Western Blot, Expressing

Journal: PLoS ONE

Article Title: Allergen-Induced Dermatitis Causes Alterations in Cutaneous Retinoid-Mediated Signaling in Mice

doi: 10.1371/journal.pone.0071244

Figure Lengend Snippet: Systemic and topical OVA sensitization results in inflammation, disturbs epidermal barrier homeostasis, and induces PPARδ target gene expression in skin.

Article Snippet: After fixation in paraformaldehyde, five-micrometer sections were incubated with a rabbit anti-mouse Fabp5 polyclonal antibody (1∶50; ProteinTech, Chicago, IL) following manufacturer’s directions after antigen retrieval ( ).

Techniques: Expressing, Binding Assay

Journal: PLoS ONE

Article Title: Allergen-Induced Dermatitis Causes Alterations in Cutaneous Retinoid-Mediated Signaling in Mice

doi: 10.1371/journal.pone.0071244

Figure Lengend Snippet: ( a ) IL-4 serum levels after systemic with or without additional topical OVA sensitization (n = 8). ( b ) ATRA levels in mouse skin determined by HPLC MS-MS method upon systemic (i.p.) and systemic plus topical (i.p.+e.c.) OVA sensitization (n = 3/group). ( c ) Ratio of Fabp5 vs. Crabp2 expression in the skin of OVA-treated mice (n = 6/group) compared to control mice (PBS i.p.). Data are presented as mean values ± SEM. Statistical significance ( p ) is based on one-way ANOVA followed by Tukey’s multiple comparison test for gene expression results and ELISA data. For HPLC MS-MS results, significance was determined using Student’s t -test.

Article Snippet: After fixation in paraformaldehyde, five-micrometer sections were incubated with a rabbit anti-mouse Fabp5 polyclonal antibody (1∶50; ProteinTech, Chicago, IL) following manufacturer’s directions after antigen retrieval ( ).

Techniques: Tandem Mass Spectroscopy, Expressing, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Allergen-Induced Dermatitis Causes Alterations in Cutaneous Retinoid-Mediated Signaling in Mice

doi: 10.1371/journal.pone.0071244

Figure Lengend Snippet: ( a ) Fabp5 protein levels in the skin of mice with allergen-induced dermatitis. 150 µg proteins were loaded per lane and beta-actin was used as control for even protein loading. ( b ) Immunohistochemical analysis of Fabp5 protein expression in five-micrometer back skin sections of OVA-sensitized mice. ( c ) ATRA-induced nuclear receptor-mediated signaling pathways depending on the predominant cellular transport protein.

Article Snippet: After fixation in paraformaldehyde, five-micrometer sections were incubated with a rabbit anti-mouse Fabp5 polyclonal antibody (1∶50; ProteinTech, Chicago, IL) following manufacturer’s directions after antigen retrieval ( ).

Techniques: Immunohistochemical staining, Expressing

Journal: iScience

Article Title: Adenosine-rich extract of Ganoderma lucidum: A safe and effective lipid-lowering substance

doi: 10.1016/j.isci.2022.105214

Figure Lengend Snippet: The levels of proteins and PTMs in the PPAR signaling pathway

Article Snippet: FABP5 Rabbit Polyclonal Antibody , proteintech , 12348-1-AP.

Techniques: Expressing, Significance Assay, Modification

Journal: iScience

Article Title: Adenosine-rich extract of Ganoderma lucidum: A safe and effective lipid-lowering substance

doi: 10.1016/j.isci.2022.105214

Figure Lengend Snippet:

Article Snippet: FABP5 Rabbit Polyclonal Antibody , proteintech , 12348-1-AP.

Techniques: Software

Journal: Cancer Biology & Therapy

Article Title: Fatty acid binding protein 5 promotes the proliferation, migration, and invasion of hepatocellular carcinoma cells by degradation of Krüppel-like factor 9 mediated by miR-889-5p via cAMP-response element binding protein

doi: 10.1080/15384047.2022.2094670

Figure Lengend Snippet: FABP5 expression was upregulated in HCC. a, expression of FABP5 in HCC samples (N = 369) and normal samples (N = 160) from the TCGA data set. b, relative expression of FABP5 mRNA in HCC tissues compared with adjacent normal tissues were detected by RT-qPCR. c, relative expression of FABP5 protein in HCC tissues and adjacent normal tissues were detected by western blotting. Representative images of FABP5 expression in 48 paired HCC tissues(t) and adjacent normal tissues (n). d, the expression of FABP5 was negatively correlated with overall survival time in HCC patients from the TCGA data set. * P < .05 and *** P < .001 as compared with the vehicle control.

Article Snippet: Nonspecific antigens were blocked in 5% skimmed milk for 1 h. Subsequently, the membranes were incubated overnight at 4°C with the different primary antibodies as follows: anti-FABP5 rabbit polyclonal antibody (Abcam, Cat. no. ab84028), anti-KLF9 rabbit polyclonal antibody (Abcepta, Cat. no. Ap16249a), anti-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 3811), anti-Phospho-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 13857), anti-AKT rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 4685), anti-Phospho-AKT rabbit monoclonal antibody (Abways, Cat. no. cy6569), anti-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 2986), anti-Phospho-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 5536), anti-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9197), anti-Phospho-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9198), anti-β-actin mouse monoclonal antibody (Santa Cruz, Dallas, Cat. no. sc-8432), and anti-GAPDH mouse monoclonal antibody (Abcam, Cat. no. ab59164).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Cancer Biology & Therapy

Article Title: Fatty acid binding protein 5 promotes the proliferation, migration, and invasion of hepatocellular carcinoma cells by degradation of Krüppel-like factor 9 mediated by miR-889-5p via cAMP-response element binding protein

doi: 10.1080/15384047.2022.2094670

Figure Lengend Snippet: Correlation between FABP5 expression and clinicopathological characteristics

Article Snippet: Nonspecific antigens were blocked in 5% skimmed milk for 1 h. Subsequently, the membranes were incubated overnight at 4°C with the different primary antibodies as follows: anti-FABP5 rabbit polyclonal antibody (Abcam, Cat. no. ab84028), anti-KLF9 rabbit polyclonal antibody (Abcepta, Cat. no. Ap16249a), anti-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 3811), anti-Phospho-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 13857), anti-AKT rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 4685), anti-Phospho-AKT rabbit monoclonal antibody (Abways, Cat. no. cy6569), anti-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 2986), anti-Phospho-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 5536), anti-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9197), anti-Phospho-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9198), anti-β-actin mouse monoclonal antibody (Santa Cruz, Dallas, Cat. no. sc-8432), and anti-GAPDH mouse monoclonal antibody (Abcam, Cat. no. ab59164).

Techniques: Expressing

Journal: Cancer Biology & Therapy

Article Title: Fatty acid binding protein 5 promotes the proliferation, migration, and invasion of hepatocellular carcinoma cells by degradation of Krüppel-like factor 9 mediated by miR-889-5p via cAMP-response element binding protein

doi: 10.1080/15384047.2022.2094670

Figure Lengend Snippet: FABP5 knockdown inhibited cell growth, migration and invasion in HCC cells. a, the mRNA and protein expression levels of FABP5 were analyzed in FABP5-knockdown HepG2 and SK-Hep-1 cells by RT-qPCR and western blotting. b, the growth curve of HCC cells was determined by CCK-8 assay. c, PI fluorescence pattern was applied for cell-cycle distribution in HCC cells transfected with shFABP5 or shNC. d, HCC cells transfected with shFABP5 or shNC were stained with Annexin-V FITC and PI to be measured apoptosis. Apoptotic cells are presented in the right-lower (Q3, early apoptosis) and right-upper (Q2, late apoptosis) quadrants of the plots. e, transwell assay revealed the migration abilities of HCC cells transfected with shFABP5 or shNC. f, transwell assay revealed the invasive abilities of HCC cells transfected with shFABP5 or shNC. Results are presented as mean ± S.E.M. (N = 3). Results were averaged from three independent experiments and presented as percentage of control levels. * p < .05, ** p < .01 and *** p < .001 as compared with the vehicle control. FITC, fluorescein isothiocyanate; PI, propidium iodide.

Article Snippet: Nonspecific antigens were blocked in 5% skimmed milk for 1 h. Subsequently, the membranes were incubated overnight at 4°C with the different primary antibodies as follows: anti-FABP5 rabbit polyclonal antibody (Abcam, Cat. no. ab84028), anti-KLF9 rabbit polyclonal antibody (Abcepta, Cat. no. Ap16249a), anti-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 3811), anti-Phospho-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 13857), anti-AKT rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 4685), anti-Phospho-AKT rabbit monoclonal antibody (Abways, Cat. no. cy6569), anti-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 2986), anti-Phospho-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 5536), anti-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9197), anti-Phospho-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9198), anti-β-actin mouse monoclonal antibody (Santa Cruz, Dallas, Cat. no. sc-8432), and anti-GAPDH mouse monoclonal antibody (Abcam, Cat. no. ab59164).

Techniques: Migration, Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Fluorescence, Transfection, Staining, Transwell Assay

Journal: Cancer Biology & Therapy

Article Title: Fatty acid binding protein 5 promotes the proliferation, migration, and invasion of hepatocellular carcinoma cells by degradation of Krüppel-like factor 9 mediated by miR-889-5p via cAMP-response element binding protein

doi: 10.1080/15384047.2022.2094670

Figure Lengend Snippet: Decreased expression of FABP5 inhibited the growth of HCC cells in vivo. a, HepG2 cells and HepG2 cells stably transfected with shFABP5 RNA or shNC were implanted into nude mice by subcutaneous injection. Tumor volume was measured each 7 days by a calliper until the end of the experiment. The tumor growth curve was drawn. b, the mice and the tumors of different groups on the 28th day after injection. c, the weight of each xenograft tumor was measured at the end of the experiment. d, the differences in FABP5 mRNA levels between the shFABP5 group and the control groups were detected by RT-qPCR. e, the protein levels of FABP5 in the shFABP5 group and the control groups were measured by western blotting. f, the tumors were analyzed by immunohistochemical staining with anti-FABP5. Original magnification: 400 × . *** p < .001 as compared with the vehicle control.

Article Snippet: Nonspecific antigens were blocked in 5% skimmed milk for 1 h. Subsequently, the membranes were incubated overnight at 4°C with the different primary antibodies as follows: anti-FABP5 rabbit polyclonal antibody (Abcam, Cat. no. ab84028), anti-KLF9 rabbit polyclonal antibody (Abcepta, Cat. no. Ap16249a), anti-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 3811), anti-Phospho-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 13857), anti-AKT rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 4685), anti-Phospho-AKT rabbit monoclonal antibody (Abways, Cat. no. cy6569), anti-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 2986), anti-Phospho-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 5536), anti-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9197), anti-Phospho-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9198), anti-β-actin mouse monoclonal antibody (Santa Cruz, Dallas, Cat. no. sc-8432), and anti-GAPDH mouse monoclonal antibody (Abcam, Cat. no. ab59164).

Techniques: Expressing, In Vivo, Stable Transfection, Transfection, Injection, Quantitative RT-PCR, Western Blot, Immunohistochemical staining, Staining

Journal: Cancer Biology & Therapy

Article Title: Fatty acid binding protein 5 promotes the proliferation, migration, and invasion of hepatocellular carcinoma cells by degradation of Krüppel-like factor 9 mediated by miR-889-5p via cAMP-response element binding protein

doi: 10.1080/15384047.2022.2094670

Figure Lengend Snippet: The expression of KLF9 was negatively correlated with FABP5. a, gene microarray analysis were applied to examine the differential expression genes in SK-Hep-1 cells before and after FABP5 knockdown (FC > |2|; P < .01). b, gene ontology (GO) enrichment analysis of differential expression genes in accordance with the biological processes, cellular component, and molecular function. c, KLF9 mRNA expression in HepG2 and SK-Hep-1 cells with FABP5 knockdown determined by RT-qPCR. d, the KLF9 expression in HCC tissue and adjacent tissues were detected by RT-qPCR. e, relative expression of KLF9 protein in HCC tissues and adjacent normal tissues were detected by Western blotting. Representative images of KLF9 expression in 48 paired HCC tissues (t) and adjacent normal tissues (n). e, the expression of KLF9 was positively correlated with overall survival time in HCC patients from the TCGA data set. F, A negative correlation was found between the protein expression level of FABP5 and KLF9 in HCC samples. *p < .05, **p < .01, and ***p < .001 as compared with the vehicle control.

Article Snippet: Nonspecific antigens were blocked in 5% skimmed milk for 1 h. Subsequently, the membranes were incubated overnight at 4°C with the different primary antibodies as follows: anti-FABP5 rabbit polyclonal antibody (Abcam, Cat. no. ab84028), anti-KLF9 rabbit polyclonal antibody (Abcepta, Cat. no. Ap16249a), anti-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 3811), anti-Phospho-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 13857), anti-AKT rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 4685), anti-Phospho-AKT rabbit monoclonal antibody (Abways, Cat. no. cy6569), anti-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 2986), anti-Phospho-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 5536), anti-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9197), anti-Phospho-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9198), anti-β-actin mouse monoclonal antibody (Santa Cruz, Dallas, Cat. no. sc-8432), and anti-GAPDH mouse monoclonal antibody (Abcam, Cat. no. ab59164).

Techniques: Expressing, Microarray, Quantitative RT-PCR, Western Blot

Journal: Cancer Biology & Therapy

Article Title: Fatty acid binding protein 5 promotes the proliferation, migration, and invasion of hepatocellular carcinoma cells by degradation of Krüppel-like factor 9 mediated by miR-889-5p via cAMP-response element binding protein

doi: 10.1080/15384047.2022.2094670

Figure Lengend Snippet: KLF9 knockdown reversed the phenotypes caused by FABP5 knockdown in HCC cells. a-b, HepG2 and SK-Hep-1 cells upon co-transfection of shFABP5 with shKLF9 showed significantly reduced KLF9 expression at the mRNA and protein levels. c, CCK-8 assay showed that shKLF9 significantly reversed the inhibition of proliferation induced by FABP5 knockdown in HCC cells. d, transwell migration assay revealed that shKLF9 significantly rescued the migratory inhibition induced FABP5 knockdown in HCC cells. e, transwell invasion assay revealed that shKLF9 significantly rescued the inhibition of invasion induced by FABP5 knockdown in HCC cells. F, KLF9 knockdown reversed the cell cycle arrest caused by FABP5 knockdown in HCC cells. PI fluorescence pattern was applied for cell-cycle distribution. G, KLF9 knockdown reversed the apoptosis caused by FABP5 knockdown in HCC cells. HepG2 and SK-Hep-1 cells were stained with Annexin-V FITC and PI to be measured apoptosis. Apoptotic cells are presented in the right-lower (Q3, early apoptosis) and right-upper (Q2, late apoptosis) quadrants of the plots. Results are presented as mean ± S.E.M. (N = 3). * p < .05, ** p < .01, and ***p < .001 as compared with the vehicle control. Results were averaged from three independent experiments and presented as percentage of control levels. FITC, fluorescein isothiocyanate; PI, propidium iodide.

Article Snippet: Nonspecific antigens were blocked in 5% skimmed milk for 1 h. Subsequently, the membranes were incubated overnight at 4°C with the different primary antibodies as follows: anti-FABP5 rabbit polyclonal antibody (Abcam, Cat. no. ab84028), anti-KLF9 rabbit polyclonal antibody (Abcepta, Cat. no. Ap16249a), anti-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 3811), anti-Phospho-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 13857), anti-AKT rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 4685), anti-Phospho-AKT rabbit monoclonal antibody (Abways, Cat. no. cy6569), anti-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 2986), anti-Phospho-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 5536), anti-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9197), anti-Phospho-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9198), anti-β-actin mouse monoclonal antibody (Santa Cruz, Dallas, Cat. no. sc-8432), and anti-GAPDH mouse monoclonal antibody (Abcam, Cat. no. ab59164).

Techniques: Cotransfection, Expressing, CCK-8 Assay, Inhibition, Transwell Migration Assay, Transwell Invasion Assay, Fluorescence, Staining

Journal: Cancer Biology & Therapy

Article Title: Fatty acid binding protein 5 promotes the proliferation, migration, and invasion of hepatocellular carcinoma cells by degradation of Krüppel-like factor 9 mediated by miR-889-5p via cAMP-response element binding protein

doi: 10.1080/15384047.2022.2094670

Figure Lengend Snippet: PI3K/AKT signaling pathway was involved in FABP5-induced KLF9 downregulation. a, KEGG pathway enrichment analysis of top 20 differentially expressed signaling pathways in FABP5 knockdown group compared with control group in SK-Hep-1 cells. b, PI3K/AKT signaling pathway was analyzed by western blotting in the indicated groups.

Article Snippet: Nonspecific antigens were blocked in 5% skimmed milk for 1 h. Subsequently, the membranes were incubated overnight at 4°C with the different primary antibodies as follows: anti-FABP5 rabbit polyclonal antibody (Abcam, Cat. no. ab84028), anti-KLF9 rabbit polyclonal antibody (Abcepta, Cat. no. Ap16249a), anti-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 3811), anti-Phospho-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 13857), anti-AKT rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 4685), anti-Phospho-AKT rabbit monoclonal antibody (Abways, Cat. no. cy6569), anti-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 2986), anti-Phospho-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 5536), anti-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9197), anti-Phospho-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9198), anti-β-actin mouse monoclonal antibody (Santa Cruz, Dallas, Cat. no. sc-8432), and anti-GAPDH mouse monoclonal antibody (Abcam, Cat. no. ab59164).

Techniques: Western Blot

Journal: Cancer Biology & Therapy

Article Title: Fatty acid binding protein 5 promotes the proliferation, migration, and invasion of hepatocellular carcinoma cells by degradation of Krüppel-like factor 9 mediated by miR-889-5p via cAMP-response element binding protein

doi: 10.1080/15384047.2022.2094670

Figure Lengend Snippet: miR-889-5p suppressed KLF9 expression by directly targeted KLF9. a, the expression of miR-889-5p was significantly decreased by FABP5 knockdown in HCC cells. b, bioinformatics analysis predicted that the 3ʹUTR sequence of KLF9 is complementary to the seed sequence of miR-889-5p.The core binding sequences of KLF9 were mutated (in red text). c, dual-luciferase reporter assay was performed to verify that miR-889-5p directly bound to the 3’-UTR sequences of KLF9 in the cells co-transfected with miR-889 mimics or NC with pGL3-KLF9-WT or pGL3-KLF9-Mut. d, relative expression of miR-889-5p in 48 HCC tissues compared with adjacent normal tissues was detected by RT-qPCR. E, miR-889-5p expression was negatively correlated with KLF9 expression and positively correlated with FABP5 expression in HCC tissues. f-g, increased expression of miR-889-5p attenuated KLF9 mRNA and protein expression of in HCC cells with shFABP5 treatment. *p < .05, **p < .01, and ***p < .001 as compared with the vehicle control.

Article Snippet: Nonspecific antigens were blocked in 5% skimmed milk for 1 h. Subsequently, the membranes were incubated overnight at 4°C with the different primary antibodies as follows: anti-FABP5 rabbit polyclonal antibody (Abcam, Cat. no. ab84028), anti-KLF9 rabbit polyclonal antibody (Abcepta, Cat. no. Ap16249a), anti-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 3811), anti-Phospho-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 13857), anti-AKT rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 4685), anti-Phospho-AKT rabbit monoclonal antibody (Abways, Cat. no. cy6569), anti-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 2986), anti-Phospho-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 5536), anti-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9197), anti-Phospho-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9198), anti-β-actin mouse monoclonal antibody (Santa Cruz, Dallas, Cat. no. sc-8432), and anti-GAPDH mouse monoclonal antibody (Abcam, Cat. no. ab59164).

Techniques: Expressing, Sequencing, Binding Assay, Luciferase, Reporter Assay, Transfection, Quantitative RT-PCR

Journal: Cancer Biology & Therapy

Article Title: Fatty acid binding protein 5 promotes the proliferation, migration, and invasion of hepatocellular carcinoma cells by degradation of Krüppel-like factor 9 mediated by miR-889-5p via cAMP-response element binding protein

doi: 10.1080/15384047.2022.2094670

Figure Lengend Snippet: FABP5 inhibits the expression of miR-889-5p by regulating CREB protein phosphorylation. a, the siRNA of NF-κB, c-Myc and CREB were transiently transferred into HepG2 and SK-Hep-1 cells. The expression of miR-889-5p was detected by RT-qPCR. b, the designed mutative model of miR-889-5p promoter. c, a luciferase reporter promoter system was employed to confirm CREB bind to miR-889-5p promoter. d, the expression of total and phosphorylated CREB protein were detected by western blotting in the indicated groups. e, CREB inhibitor KG-501 reverse the effect of FABP5 overexpression on miR-889-5p expression in HCC cells. **p < .01 and ***p < .001 as compared with the vehicle control. e, the scheme of the mechanism by which FABP5 affects HCC tumorigenesis. FABP5 improves CREB protein phosphorylation to upregulate the miR-889-5p expression by CREB binding to the miR-889-5p promoter region, whereby leading to downregulation of KLF9 by miR-889-5p binding to the 3ʹ-UTR of the KLF9 mRNA, potentiating the PI3K/AKT signaling pathway and promoting the proliferation, migration, and invasion of HCC cells.

Article Snippet: Nonspecific antigens were blocked in 5% skimmed milk for 1 h. Subsequently, the membranes were incubated overnight at 4°C with the different primary antibodies as follows: anti-FABP5 rabbit polyclonal antibody (Abcam, Cat. no. ab84028), anti-KLF9 rabbit polyclonal antibody (Abcepta, Cat. no. Ap16249a), anti-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 3811), anti-Phospho-PI3K rabbit polyclonal antibody (Cell Signaling Technology, Cat. no. 13857), anti-AKT rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 4685), anti-Phospho-AKT rabbit monoclonal antibody (Abways, Cat. no. cy6569), anti-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 2986), anti-Phospho-mTOR rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 5536), anti-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9197), anti-Phospho-CREB rabbit monoclonal antibody (Cell Signaling Technology, Cat. no. 9198), anti-β-actin mouse monoclonal antibody (Santa Cruz, Dallas, Cat. no. sc-8432), and anti-GAPDH mouse monoclonal antibody (Abcam, Cat. no. ab59164).

Techniques: Expressing, Quantitative RT-PCR, Luciferase, Western Blot, Over Expression, Binding Assay, Migration