Journal: Cell Regeneration

Article Title: Identification of feature genes in intestinal epithelial cell types

doi: 10.1186/s13619-024-00208-8

Figure Lengend Snippet: The expression of specific transcription factors, membrane-associated proteins and cell markers in human small and large intestine. A Immunoblot of protein FOXM1, MYBL2 and UBE2C in villus and crypt derived from mouse small intestine. B FOXM1 and KI67 immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. C UBE2C immunostaining of large intestine derived from human specimens. Scale bars: 10 μm. D MYBL2 and KI67 immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. E Immunoblot of protein ANPEP, SLC12A2, IGF1R and SLC16A1 in villus and crypt derived from mouse small intestine. F SLC12A2 immunostaining of small intestine derived from human specimens. Scale bars: 50 μm. G SLC16A1 and E-Cadherin immunostaining of small intestine derived from human specimens. Scale bars: 50 μm. H ANPEP and E-Cadherin immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. I IGF1R immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. J Immunoblot of protein HSPD1, CPE, FABP5 and C1QBP in villus and crypt derived from mouse small intestine. K CPE and Chromogranin A immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. L HSPD1 and E-Cadherin immunostaining of small intestine derived from human specimens. Scale bars: 50 μm. M FABP5 and Chromogranin A immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. N C1QBP and E-Cadherin immunostaining of small intestine derived from human specimens. Scale bars: 50 μm

Article Snippet: Mouse anti-Ki67 (1:200; 9449, CST), mouse anti-E-cadherin (1:1000; 610,182, BD Biosciences), mouse anti-chromogranin A (1:200; sc-393941, Santa Cruz), rabbit anti-Foxm1 (1:100; ab207298, Abcam), rabbit anti-Ube2c (1:50; 12,134–2-AP, Proteintech), rabbit anti-Mybl2 (1:100; ab76009, Abcam), rabbit anti-Anpep (1:50; 14,553–1-AP, Proteintech), rabbit anti-Slc12a2 (1:50; 13,884–1-AP, Proteintech), rabbit anti-Slc16a1 (1:50; 20,139–1-AP, Proteintech), rabbit anti-Igf1r (1:100; ab182408, Abcam), rabbit anti-Cpe (1:50; 13,710–1-AP, Proteintech), rabbit anti-Hspd1 (1:50; 15,282–1-AP, Proteintech), rabbit anti-C1qbp (1:50; 24,474–1-AP, Proteintech), rabbit anti-Fabp5 (1:100; ab255276, Abcam).

Techniques: Expressing, Membrane, Western Blot, Derivative Assay, Immunostaining

Journal: bioRxiv

Article Title: Lysosomes cell autonomously regulate myeloid cell states and immune responses

doi: 10.1101/2024.11.11.623074

Figure Lengend Snippet: Grn KO myeloid cells display an MG8-like signature in vitro . (a) Average expression of top 150 regulated genes [from Grn KO primary microglia (pMG) in vitro ] in each microglia subcluster from aged Grn KO microglia scRNA-seq. (b) Heatmap of relative expression of top 175 MG8 markers (from aged Grn KO microglia scRNA-seq) in WT and Grn KO pMG in vitro . (c) Volcano plot displaying differential expression between Grn KO and WT pMG. Top MG8 marker genes are highlighted as triangles. (d,e) Western blots and quantification of MG8 markers (GPNMB, FABP5, and total CatB) and LPL (not expressed by MG8) from protein lysates of WT and Grn KO mouse primary microglia (pMG) (n=5 biological replicates/genotype). (f) Average expression of top 150 upregulated genes [from Grn KO bone marrow-derived macrophages (BMDMs) in vitro ] in each microglia subcluster from aged Grn KO microglia scRNA-seq. (g) Heatmap of relative expression of top 175 MG8 markers (from aged Grn KO microglia scRNA-seq) in WT and Grn KO BMDMs in vitro . (h) Volcano plot displaying differential expression between Grn KO and WT BMDMs. Top MG8 marker genes are highlighted as triangles. (i) Western blots and quantification of MG8 markers (GPNMB, FABP5, and total CatB) and LPL (not expressed by MG8) from protein lysates of WT and Grn KO mouse BMDMs (n=3 biological replicates/genotype). Data presented in (e,j) are mean±s.e.m. Student’s t-tests were performed to compare across genotypes in (e,j). * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001.

Article Snippet: The following primary antibodies were used: sheep anti-PGRN (R&D Systems; Cat# 2557; 0.5μg/mL), rabbit anti-GPNMB (Cell Signaling Technology; Cat# 90205; 1:500), rabbit anti-FABP5 (Cell Signaling Technology; Cat# 39926; 1:500), rabbit anti-CatB (Cell Signaling Technology; Cat# 31718; 1:500), rabbit anti-LPL (Biotechne; Cat# AF7197; 1μg/mL), mouse anti-Vinculin (Sigma-Aldrich; Cat# V9264; 1:1000), mouse anti-β-Actin (Sigma-Aldrich; Cat# A2228; 1:10000), mouse anti-LC3 (Cell Signaling Technology; Cat# 83506; 1:500), mouse anti-p62 (Biotechne; Cat# MAB8028; 2μg/mL), mouse anti-AMPKα (Cell Signaling Technology; Cat# 2793; 1:500), rabbit anti-phospho-AMPKα (T172) (Cell Signaling Technology; Cat# 2535; 1:500), mouse anti- GAPDH (Abcam; Cat# ab8245; 1:2000), rabbit anti-ATP6V1A (Cell Signaling Technology; Cat# 39517; 1:500), rabbit anti-NPC1 (Abcam; Cat# ab134113; 1:500), rabbit anti-VPS34 (Cell Signaling Technology; Cat# 4263; 1:500), rabbit anti-CatD (Cell Signaling Technology; Cat# 88239; 1:500), sheep anti-TREM2 (Biotechne; Cat# AF1729; 0.1μg/mL), rabbit anti-ATG7 (Cell Signaling Technology; Cat# 8558; 1:500), rabbit anti-ATG14 (Cell Signaling Technology; Cat# 96752; 1:500), and rabbit anti-UVRAG (Abcam; Cat# ab313627; 1:500).

Techniques: In Vitro, Expressing, Marker, Western Blot, Derivative Assay

Journal: iScience

Article Title: OBP2A regulates epidermal barrier function and protects against cytotoxic small hydrophobic molecules

doi: 10.1016/j.isci.2024.111093

Figure Lengend Snippet:

Article Snippet: FABP5 (D1A7T) Rabbit mAb , Cell Signaling Technology , 39926.

Techniques: Imaging, Recombinant, Transfection, Saline, Sterility, Cell Culture, Protease Inhibitor, DC Protein Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Quantitation Assay, Fluorescence, Sequencing, Software

Journal: International Journal of Oncology

Article Title: FABP5 can substitute for androgen receptor in malignant progression of prostate cancer cells

doi: 10.3892/ijo.2023.5606

Figure Lengend Snippet: Proteins detected by western blot and antibodies used.

Article Snippet: FABP5 , Monoclonal Rabbit Anti-human FABP5 (1:500; Hycult; cat. no. HP-9030) , Swine polyclonal anti-rabbit Immunoglobulin/HRP (Dako; Agilent Technologies, Inc.; 1:20,000; cat no: P039901-2).

Techniques: Western Blot, Marker

Journal: International Journal of Oncology

Article Title: FABP5 can substitute for androgen receptor in malignant progression of prostate cancer cells

doi: 10.3892/ijo.2023.5606

Figure Lengend Snippet: The guide RNAs, PAM sequences, and the genomic locations of the KO genes.

Article Snippet: FABP5 , Monoclonal Rabbit Anti-human FABP5 (1:500; Hycult; cat. no. HP-9030) , Swine polyclonal anti-rabbit Immunoglobulin/HRP (Dako; Agilent Technologies, Inc.; 1:20,000; cat no: P039901-2).

Techniques:

Journal: International Journal of Oncology

Article Title: FABP5 can substitute for androgen receptor in malignant progression of prostate cancer cells

doi: 10.3892/ijo.2023.5606

Figure Lengend Snippet: Western blot analysis of some key proteins associated with the FABP5 pathway in benign and malignant prostate cells, and western blot analysis was combined with DNA-sequencing analysis to verify the successful gene KO cell clones. An anti-β-actin antibody was incubated with each blot to standardize the immunological responses. Quantitative analysis was performed by densitometrical scanning of the area and peak of each band on the blot. The protein relative levels were reproduced independently three times, and the differences were assessed by Student's t-test for results with 2 datasets, and one-way ANOVA test for results with 3 or more datasets. Dunnett's post hoc test was utilized for multiple comparisons following the ANOVA. (A) Relative expression of FABP5 pathway-associated proteins in benign and malignant prostate epithelial cells is shown. a) Western blot analysis of FABP5 in benign PNT2 cells, the moderately malignant 22RV1 cells, and the highly malignant prostate carcinoma cell lines, DU145 and PC3M. b) Relative levels of FABP5 protein in PNT2, 22RV1, DU145 and PC3M cells. The level of FABP5 in 22RV1 cells was set at '1', and the levels in DU145 and PC3M were obtained by comparing with that of 22RV1. c) Western blot detection of PPARγ in PNT2, 22RV1, DU145 and PC3M cells. d) Relative levels of PPARγ in PNT2, 22RV1, DU145, and PC3M cells. The level in PNT2 cells was set at '1', and the levels in the other cell lines were obtained by comparing with that of PNT2. e) Western blot analysis of pPPARγ in PNT2, 22RV1, DU145 and PC3M cells. f) Relative levels of pPPARγ in PNT2, 22RV1, DU145, and PC3M cells. The level in 22RV1 cells was set at '1', and levels in DU145 and PC3M were obtained by comparing with that of 22RV1. g) Western blot analysis of ARFL protein in PNT2, 22RV1, DU145 and PC3M cells. h) Relative levels of ARFL in prostate cell lines. The level of ARFL in 22RV1 cells was set at '1', and levels in the other cell lines were not detectable. i) Western blot analysis of VEGF protein in PNT2, 22RV1, DU145 and PC3M cells. j) Relative levels of VEGF; the level in PNT2 was set at '1', and levels in the other cell lines were obtained by comparing with that of PNT2. (B) Western blot and DNA sequencing analyses were performed to verify successful gene suppression in KO cell clones. a) Western blots of FABP5 protein in 22RV1 cells and its derivative FABP5 -KO clones. b) Relative levels of FABP5 in 22RV1 cells and in its derivative FABP5 -KO clones. The level of FABP5 in 22RV1 cells was set at '1', and levels in different clones were obtained by comparing with that in 22RV1. c) Further Western blots of FABP5 protein in 22RV1 cells and in the successful FABP5 -KO clone C3. d) DNA sequence analysis revealed the location where the guide RNA-induced mutation occurred (depicted by the black arrow). e) Western blot analysis of FABP5 protein in DU145 cells and in its derivative FABP5 -KO clones. f) Relative levels of FABP5 in DU145 and in the different FABP5 -KO clones. The level of FABP5 in DU145 was set at '1', and the levels in the different clones were obtained by comparing with that in DU145. g) Western blot analysis of FABP5 protein in DU145 cells and in a selected FABP5 -KO clone (C2). h) DNA sequence analysis revealed the location where the guide RNA-induced mutation occurred (depicted by the black arrow). i) Western blot detection of FABP5 expression in PC3M and in different FABP5 -KO clones. j) Relative levels of FABP5 in PC3M and in its different FABP5 -KO clones. The level of FABP5 in PC3M was set at '1', and the levels in its different FABP5 -KO clones were obtained by comparing with that in PC3M. k) Western blot analysis of FABP5 protein in PC3M and in its selected FABP5 -KO clone, C6. l) DNA sequence analysis revealed the location where the guide RNA-induced mutation occurred (depicted by the black arrow). m) Western blot analysis of AR protein in 22RV1 cells and in its different AR -KO clones. n) Relative levels of AR in 22RV1 cells and in its AR -KO clones. The level of AR in 22RV1 was set at '1', and the levels of AR in different clones were obtained by comparing with that in 22RV1. o) Western blot analysis of AR expression in 22RV1 cells and in its AR -KO clones. p) Relative levels of AR in 22RV1 and in its AR -KO clones. The level of AR in 22RV1 was set at '1', and the levels of AR in different clones were obtained by comparing with that in 22RV1 cells. *** P<0.0001 and **** P<0.00001. KO, knockout; FABP5, fatty acid-binding protein 5; AR, androgen receptor; PPARγ, peroxisome proliferator-activated receptor-γ; VEGF, vascular endothelium growth factor; pPPARγ, phosphorylated PPARγ; ns, not statistically significant.

Article Snippet: FABP5 , Monoclonal Rabbit Anti-human FABP5 (1:500; Hycult; cat. no. HP-9030) , Swine polyclonal anti-rabbit Immunoglobulin/HRP (Dako; Agilent Technologies, Inc.; 1:20,000; cat no: P039901-2).

Techniques: Western Blot, DNA Sequencing, Clone Assay, Incubation, Expressing, Sequencing, Mutagenesis, Knock-Out, Binding Assay

Journal: International Journal of Oncology

Article Title: FABP5 can substitute for androgen receptor in malignant progression of prostate cancer cells

doi: 10.3892/ijo.2023.5606

Figure Lengend Snippet: Effect of FABP5 -KO on malignant characteristics of 22RV1 cells and on the levels of downstream proteins of the FABP5 signaling pathway. (A) Microscopical appearances of 22RV1-FABP5-KO cells and their parental 22RV1 cells. (B) Effect of FABP5 -KO on proliferation of 22RV1 cells. C) Effect of FABP5 -KO on invasion of 22RV1 cells. (D) Quantitative assessment of the numbers of invasive cells. (E) Effect of FABP5 -KO on anchorage-independent growth of 22RV1 cells. (F) Quantitative assessment on cell colony numbers formed in soft agar. (G) Effect of FABP5 -KO on motility of 22RV1 cells. (H) Quantitative assessment of the average wound width of the space of the gap ( μ m) at different times is shown. (I) Western blot analysis of PPARγ1 and PPARγ2 in 22RV1 and in 22RV1-FABP5-KO cells. (J) Relative levels of PPARγ1 and PPARγ2. The levels of PPARγ1 and PPARγ2 in 22RV1 cells were each set at '1', and their levels in 22RV1-FABP5-KO cells were obtained by comparing with those in 22RV1. (K) Western blot analysis of pPPARγ1 and pPPARγ2 in 22RV1 and in 22RV1-FABP5-KO cells. (L) Relative levels of pPPARγ1 and pPPARγ2. The levels of pPPARγ1 and pPPARγ2 in 22RV1 were each set at '1', and those in 22RV1-FABP5-KO cells were obtained by comparing with those in 22RV1. (M) Western blot analysis of VEGF in 22RV1 and in 22RV1-FABP5-KO cells. (N) Relative levels of VEGF protein. The level of VEGF in 22RV1 cells was set at '1', and the level of VEGF in 22RV1-FABP5-KO cells was obtained by comparing with that in 22RV1. (O) Western blot analysis of ARFL and ARV7 in 22RV1 and in 22RV1-FABP5-KO cells. (P) Relative levels of ARFL and ARV7. Levels of ARFL and ARV7 in 22RV1 cells were set at '1', and the levels of ARFL and ARV7 in 22RV1-FABP5-KO cells were obtained by comparing with those in 22RV1. The protein relative levels were reproduced independently three times, and the differences were assessed by Student's t-test. Results were considered significant when P<0.05. ** P<0.001, *** P<0.0001 and **** P<0.00001. KO, knockout; FABP5, fatty acid-binding protein 5; AR, androgen receptor; ARFL, full-length AR; ARV7, AR splicing variant 7; PPARγ, peroxisome proliferator-activated receptor-γ; pPPARγ, phosphorylated PPARγ; ns, not statistically significant.

Article Snippet: FABP5 , Monoclonal Rabbit Anti-human FABP5 (1:500; Hycult; cat. no. HP-9030) , Swine polyclonal anti-rabbit Immunoglobulin/HRP (Dako; Agilent Technologies, Inc.; 1:20,000; cat no: P039901-2).

Techniques: Western Blot, Knock-Out, Binding Assay, Variant Assay

Journal: International Journal of Oncology

Article Title: FABP5 can substitute for androgen receptor in malignant progression of prostate cancer cells

doi: 10.3892/ijo.2023.5606

Figure Lengend Snippet: Effect of AR -KO on the malignant characteristics of 22RV1 cells and on levels of the down-stream proteins of the FABP5 signalling pathway. (A) Microscopical appearances of 22RV1 and 22RV1-AR-KO. (B) Effect of AR -KO on proliferation of 22RV1 and 22RV1-AR-KO cells. (C) Effect of AR -KO on 22RV1 cell invasion. (D) Average number of invasive cells from 22RV1 and 22RV1-AR-KO cells. (E) Effect of AR knockout on anchorage-independent growth of 22RV1 cells. (F) Relative numbers of colonies formed in soft agar by 22RV1 and 22RV1-AR-KO cells. (G) Effect of AR -KO on gap closure of 22RV1 and 22RV1-AR-KO cells. (H) Quantitative assessment of average sizes of the wound space or gap in 22RV1 and 22RV1-AR-KO cells in μ m at different times. (I) Western blot analysis of PPARγ1 and PPARγ2 in 22RV1 and 22RV1-AR-KO cells. (J) Relative levels of PPARγ1 and PPARγ2 in 22RV1 and 22RV1-AR-KO cells. (K) Western blot analysis of pPPARγ1 and pPPARγ2 in 22RV1 and 22RV1-AR-KO cells. (L) Quantitative assessment of levels of pPPARγ1 and pPPARγ2 proteins. Levels of pPPARγ1 and pPPARγ2 in 22RV1 were set at '1', and the levels in 22RV1AR-KO cells were obtained by comparing with those in 22RV1. (M) Western blot analysis of VEGF in 22RV1 and 22RV1-AR-KO cells. (N) Relative levels of VEGF in 22RV1 and 22RV1-AR-KO cells. The level of VEGF in 22RV1 cells was set at '1', and that in 22RV1-AR-KO cells was obtained by comparing with that in 22RV1. (O) Western blot detection of FABP5 protein in 22RV1 and 22RV1-AR-KO cells (no quantification was made, since the level in 22RV1-FABP5-KO cells was negligible). The protein relative levels were reproduced independently three times, and the differences were assessed by Student's t-test. * P<0.05, *** P<0.0001 and **** P<0.00001. KO, knockout; FABP5, fatty acid-binding protein 5; AR, androgen receptor; PPARγ, peroxisome proliferator-activated receptor-γ; pPPARγ, phosphorylated PPARγ; VEGF, vascular endothelium growth factor; ns, not statistically significant.

Article Snippet: FABP5 , Monoclonal Rabbit Anti-human FABP5 (1:500; Hycult; cat. no. HP-9030) , Swine polyclonal anti-rabbit Immunoglobulin/HRP (Dako; Agilent Technologies, Inc.; 1:20,000; cat no: P039901-2).

Techniques: Knock-Out, Western Blot, Binding Assay

Journal: International Journal of Oncology

Article Title: FABP5 can substitute for androgen receptor in malignant progression of prostate cancer cells

doi: 10.3892/ijo.2023.5606

Figure Lengend Snippet: Effect of FABP5 -KO on the malignant characteristics of DU145 and on protein levels of the downstream proteins of the FABP5 pathway. (A) Microscopical appearances revealed morphological difference between DU145 and DU145-FABP5-KO cells. (B) Effect of FABP5 -KO on the proliferation of DU145 cells. (C) Effect of FABP5 -KO on DU145 cell invasion was assessed by an invasion assay, and (D) the average number of invasive cells from DU145 and DU145-FABP5-KO cells were recorded. (E) Anchorage-independent growth of DU145 and DU145-FABP5-KO cells, and (F) their average colony numbers formed in soft agar. (G) Effect of FABP5 -KO on migration was assessed at different times in a gap closure assay. (H) The quantitative assessment of the average size of wound space gap at different times. (I) Western blot analysis of PPARγ1 and PPARγ2 proteins. (J) Relative levels of PPARγ1 and PPARγ2 in DU145 and DU145-FABP5-KO cells. (K) Western blots were performed for pPPARγ1 and pPPARγ2 in DU145 and DU145-FABP5-KO cells. (L) Relative levels of pPPARγ1 and pPPARγ2. Levels of pPPARγ1 and pPPARγ2 in DU145 cells were each set at '1', and the levels in DU145-FABP5-KO cells were obtained by comparing with those in DU145. (M) Western blot analysis of VEGF protein in DU145 and DU145-FABP5-KO cells. (N) Relative levels of VEGF in DU145 and DU145-FABP5-KO cells; the level of VEGF in DU145 cells was set at '1', and the level in DU145-FABP5-KO cells was obtained by comparing with that in DU145. (O) Western blot analysis of ARFL and ARV7 proteins in DU145 and DU145-FABP5-KO cells. Since neither of the proteins were detectable in either cell line, no quantification was possible. The relative protein levels were reproduced independently three times, and the differences were assessed by Student's t-test. *** P<0.0001 and **** P<0.00001. KO, knockout; FABP5, fatty acid-binding protein 5; AR, androgen receptor; ARFL, full-length AR; ARV7, AR splicing variant 7; PPARγ, peroxisome proliferator-activated receptor-γ; pPPARγ, phosphorylated PPARγ; VEGF, vascular endothelium growth factor; ns, not statistically significant.

Article Snippet: FABP5 , Monoclonal Rabbit Anti-human FABP5 (1:500; Hycult; cat. no. HP-9030) , Swine polyclonal anti-rabbit Immunoglobulin/HRP (Dako; Agilent Technologies, Inc.; 1:20,000; cat no: P039901-2).

Techniques: Invasion Assay, Migration, Western Blot, Knock-Out, Binding Assay, Variant Assay

Journal: International Journal of Oncology

Article Title: FABP5 can substitute for androgen receptor in malignant progression of prostate cancer cells

doi: 10.3892/ijo.2023.5606

Figure Lengend Snippet: Effect of FABP5 -KO on malignant characteristics of PC3M and on levels of the downstream proteins of the FABP5 signalling pathway. (A) Microscopical images of PC3M and PC3M-FABP5-KO cells. (B) Proliferation rates of PC3M and PC3M-FABP5-KO cells over the course of the 6-day experimental period. (C) Images of invasive cells from PC3M and PC3M-FABP5-KO at the end of the 6-day experimental period. (D) The average numbers of invasive cells from PC3M and PC3M-FABP5-KO cells. (E) Images of colonies in soft agar formed by PC3M and PC3M-FABP5-KO cells. (F) Average numbers of colonies in soft agar formed by PC3M and PC3M-FABP5-KO cells. (G) Images of wounds in PC3M and PC3M-FABP5-KO cells at the different times. (H) Quantitative assessment of the average size of wound space gap ( μ m) in PC3M and PC3M-FABP5-KO cells. (I) Western blot analysis of PPARγ1 and PPARγ2 in PC3M and PC3M-FABP5-KO cells. (J) Relative levels of PPARγ1 and PPARγ2. Levels of PPARγ1 and PPARγ2 in PC3M were each set at '1', and the levels in PC3M-FABP5-KO cells were obtained by comparing with those in PC3M. (K) Western blots of pPPARγ1 and pPPARγ2 in PC3M and PC3M-FABP5-KO cells. (L) Relative levels of pPPARγ1 and pPPARγ2. Levels of pPPARγ1 and pPPARγ2 in PC3M cells were each set at '1', and the levels in PC3M-FABP5-KO cells were obtained by comparing with those in PC3M. (M) Western blot analysis of VEGF in PC3M and PC3M-FABP5-KO cells was performed. (N) The relative level of VEGF. The level of VEGF in PC3M cells was set at '1', and the level of VEGF in the PC3M-FABP5-KO cells was obtained by comparing with that in the PC3M cells. (O) Western blot analysis of ARFL and ARV7 in the PC3M and PC3M-FABP5-KO cells. Since only ARFL was present, and no ARV7 was detected, no quantification of the data was possible. The relative protein levels were reproduced independently three times, and the differences were assessed by Student's t-test. ** P<0.001, *** P<0.0001 and **** P<0.00001. KO, knockout; FABP5, fatty acid-binding protein 5; AR, androgen receptor; ARFL, full-length AR; ARV7, AR splicing variant 7; PPARγ, peroxisome proliferator-activated receptor-γ; pPPARγ, phosphorylated PPARγ; VEGF, vascular endothelium growth factor; ns, not statistically significant.

Article Snippet: FABP5 , Monoclonal Rabbit Anti-human FABP5 (1:500; Hycult; cat. no. HP-9030) , Swine polyclonal anti-rabbit Immunoglobulin/HRP (Dako; Agilent Technologies, Inc.; 1:20,000; cat no: P039901-2).

Techniques: Western Blot, Knock-Out, Binding Assay, Variant Assay

Journal: International Journal of Oncology

Article Title: FABP5 can substitute for androgen receptor in malignant progression of prostate cancer cells

doi: 10.3892/ijo.2023.5606

Figure Lengend Snippet: Identification of DEGs between the parental cells and their FABP5 - or AR -KO derivatives and the relevant pathways. (A) Top up (red)- or down (green)-regulated 40 DEGs identified from analysis of the microarray heatmaps of 22RV1 and 22RV1-FABP5-KO cells. Red color represents 'up'. (B) Top 30 pathways associated with DEGs with the 22RV1-FABP5-KO cells were revealed by the enrichment bar chart method. (C) Top up (red)- or down (green)-regulated 40 DEGs from analysis of microarray heatmaps of 22RV1 and 22RV1-AR-KO cells. (D) The top 30 pathways associated with the DEGs in 22RV1-AR-KO cells were identified by the enrichment bar chart method. (E) Top up (red)- or down (green)-regulated 40 DEGs identified by analysis of microarray heatmaps of the DU145 and DU145-FABP5-KO cells. (F) Enrichment bar chart, revealing the top 30 pathways associated with DEGs of DU145-FABP5-KO cells. DEGs, differentially expressed genes; KO, knockout; FABP5, fatty acid-binding protein 5; AR, androgen receptor.

Article Snippet: FABP5 , Monoclonal Rabbit Anti-human FABP5 (1:500; Hycult; cat. no. HP-9030) , Swine polyclonal anti-rabbit Immunoglobulin/HRP (Dako; Agilent Technologies, Inc.; 1:20,000; cat no: P039901-2).

Techniques: Microarray, Knock-Out, Binding Assay

Journal: International Journal of Oncology

Article Title: FABP5 can substitute for androgen receptor in malignant progression of prostate cancer cells

doi: 10.3892/ijo.2023.5606

Figure Lengend Snippet: Identification and verification of the six most prominent DEGs between 22RV1 cells and their FABP5 - or AR -KO derivatives, and between DU145 and DU145-FABP5-KO cells. (A) The top 6 DEGs between 22RV1 and 22RV1-FABP5-KO cells, or between 22RV1 and 22RV1-AR-KO cells (cells are represented by different colors), as identified by heatmap analysis. (B) The top 6 DEGs between DU145 and DU145-FABP5-KO cells identified by heatmap analysis. (C) Western blot verification of the top 3 up- and down-regulated DEGs between 22RV1 and 22RV1-FABP5-KO cells. (D) Relative levels of the top 3 up- and down-regulated DEGs between 22RV1 and 22RV1-FABP5-KO cells. (E) Western blot verification of the top 3 up- and down-regulated DEGs between 22RV1 and 22RV1-AR-KO cells. (F) Relative levels of the top 3 up- and down-regulated DEGs between 22RV1 and 22RV1-AR-KO cells. (G) Western blot verification of the top 3 up- and down-regulated DEGs between DU145 and DU145-FABP5-KO cells. (H) Relative levels of the top 3 up- and down-regulated DEGs between DU145 and DU145-FABP5-KO cells. The relative protein levels were reproduced independently three times, and the differences were assessed by Student's t-test. *** P<0.0001 and **** P<0.00001. DEGs, differentially expressed genes; KO, knockout; FABP5, fatty acid-binding protein 5; AR, androgen receptor; ns, not statistically significant.

Article Snippet: FABP5 , Monoclonal Rabbit Anti-human FABP5 (1:500; Hycult; cat. no. HP-9030) , Swine polyclonal anti-rabbit Immunoglobulin/HRP (Dako; Agilent Technologies, Inc.; 1:20,000; cat no: P039901-2).

Techniques: Western Blot, Knock-Out, Binding Assay

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