Review

mouse anti lamb2 (Santa Cruz Biotechnology)

Santa Cruz Biotechnology is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Santa Cruz Biotechnology mouse anti lamb2
Mouse Anti Lamb2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 71 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse anti lamb2/product/Santa Cruz Biotechnology
Average 93 stars, based on 71 article reviews

mouse anti lamb2 - by Bioz Stars, 2026-05

93/100 stars

Images

Image Search Results

Staining for the podocyte extracellular matrix proteins laminin-β2 (LAMB2), agrin, and collagen type IV-α2 (COL4A4) are increased in parietal epithelial cells (PECs) in experimental focal segmental glomerulosclerosis (FSGS). A−C: mice at baseline. LAMB2 (A), agrin (B), and COL4A4 (C) immunofluorescent staining (green) was limited to the glomerular basement membrane (yellow arrowheads). Nuclei were stained with DAPI (blue). D−F: experimental FSGS in mice. De novo immunofluorescent staining for LAMB2 (D), agrin (E), and COL4A4 (F) was detected in PECs at day 28 (D28) FSGS (white arrows). G−I: quantification. The percentage of glomerular cross-sections with de novo staining for LAMB2 (G), agrin (H), and COL4A4 (I) increased at day 28 FSGS compared with baseline. The number of mice quantitated is shown below each graph. J−L: graphs of quantification of fluorescence intensity. Fluorescence intensity was increased for LAMB2 (J), agrin (K), and COL4A4 (L) in abnormal glomeruli in FSGS. Original magnification: ×400. Scale bars = 20 μm.

Journal: American Journal of Physiology - Renal Physiology

Article Title: Differential expression of parietal epithelial cell and podocyte extracellular matrix proteins in focal segmental glomerulosclerosis and diabetic nephropathy

doi: 10.1152/ajprenal.00266.2019

Figure Lengend Snippet: Staining for the podocyte extracellular matrix proteins laminin-β2 (LAMB2), agrin, and collagen type IV-α2 (COL4A4) are increased in parietal epithelial cells (PECs) in experimental focal segmental glomerulosclerosis (FSGS). A−C: mice at baseline. LAMB2 (A), agrin (B), and COL4A4 (C) immunofluorescent staining (green) was limited to the glomerular basement membrane (yellow arrowheads). Nuclei were stained with DAPI (blue). D−F: experimental FSGS in mice. De novo immunofluorescent staining for LAMB2 (D), agrin (E), and COL4A4 (F) was detected in PECs at day 28 (D28) FSGS (white arrows). G−I: quantification. The percentage of glomerular cross-sections with de novo staining for LAMB2 (G), agrin (H), and COL4A4 (I) increased at day 28 FSGS compared with baseline. The number of mice quantitated is shown below each graph. J−L: graphs of quantification of fluorescence intensity. Fluorescence intensity was increased for LAMB2 (J), agrin (K), and COL4A4 (L) in abnormal glomeruli in FSGS. Original magnification: ×400. Scale bars = 20 μm.

Article Snippet: The C4 (mouse anti-human LAMB2) antibody was obtained from the Developmental Studies Hybridoma Bank (deposited by J.R. Sanes), developed under the auspices of Eunice Kennedy Shriver National Institute of Child Health and Human Development and maintained by the Department of Biology, The University of Iowa, Iowa City, IA.

Techniques: Staining, Membrane, Fluorescence

Double staining confirmed laminin-β2 (LAMB2) expression in parietal epithelial cells (PECs) in mice with experimental focal segmental glomerulosclerosis (FSGS). A and B: double staining for nephrin (red) and LAMB2 (green). A and A3: a normal mouse glomerulus. The merged image shows staining for LAMB2 (green) in the glomerular basement membrane distribution, with nephrin staining (red) alongside. A1 and A2: staining for nephrin (A1) and LAMB2 (A2) showing their absence along Bowman’s capsule. B and B3: FSGS. The merged image shows de novo LAMB2 but not nephrin staining along Bowman’s capsule. B1 and B2: nephrin (B1) and LAMB2 (B2) staining. Higher magnification of the inset in B showed no staining for nephrin (B4) along Bowman’s capsule in areas staining for LAMB2 (white arrows) (B5). Their merged image is shown in B6. C and D: double staining for β-Gal (red) and LAMB2 (green). C and C3: a normal glomerulus. The merged image shows that there was no overlap of LAMB2 staining in the glomerular basement membrane distribution and β-Gal, which permanently labels PECs. C1 and C2: β-Gal and LAMB2. D and D3: FSGS. The merged image shows de novo staining for LAMB2 in a Bowman’s capsule distribution (D1 and D2). Higher power magnification of the inset D shows that β-Gal (yellow arrows) (D4) is lined by LAMB2 (white arrows) (D5), as easily seen in the merged image (D6). Original magnification: ×400. Scale bars = 20 μm.

Journal: American Journal of Physiology - Renal Physiology

Article Title: Differential expression of parietal epithelial cell and podocyte extracellular matrix proteins in focal segmental glomerulosclerosis and diabetic nephropathy

doi: 10.1152/ajprenal.00266.2019

Figure Lengend Snippet: Double staining confirmed laminin-β2 (LAMB2) expression in parietal epithelial cells (PECs) in mice with experimental focal segmental glomerulosclerosis (FSGS). A and B: double staining for nephrin (red) and LAMB2 (green). A and A3: a normal mouse glomerulus. The merged image shows staining for LAMB2 (green) in the glomerular basement membrane distribution, with nephrin staining (red) alongside. A1 and A2: staining for nephrin (A1) and LAMB2 (A2) showing their absence along Bowman’s capsule. B and B3: FSGS. The merged image shows de novo LAMB2 but not nephrin staining along Bowman’s capsule. B1 and B2: nephrin (B1) and LAMB2 (B2) staining. Higher magnification of the inset in B showed no staining for nephrin (B4) along Bowman’s capsule in areas staining for LAMB2 (white arrows) (B5). Their merged image is shown in B6. C and D: double staining for β-Gal (red) and LAMB2 (green). C and C3: a normal glomerulus. The merged image shows that there was no overlap of LAMB2 staining in the glomerular basement membrane distribution and β-Gal, which permanently labels PECs. C1 and C2: β-Gal and LAMB2. D and D3: FSGS. The merged image shows de novo staining for LAMB2 in a Bowman’s capsule distribution (D1 and D2). Higher power magnification of the inset D shows that β-Gal (yellow arrows) (D4) is lined by LAMB2 (white arrows) (D5), as easily seen in the merged image (D6). Original magnification: ×400. Scale bars = 20 μm.

Techniques: Double Staining, Expressing, Staining, Membrane

The podocyte-specific extracellular matrix proteins laminin-β2 (LAMB2), agrin, and collagen type IV-α4 (COL4A4) increased in parietal epithelial cells (PECs) in human focal segmental glomerulosclerosis (FSGS). A–C: LAMB2 (green) and DAPI (blue). A: LAMB2 localized to the glomerular basement membrane (GBM) in the normal human kidney. B and C: two representative glomeruli from different patients with FSGS showing LAMB2 staining along Bowman’s capsule. B′ and C′: higher magnification of the insets showing LAMB2 staining (white arrows) along Bowman’s capsule. D–F: agrin (green) and DAPI. D: agrin localized to the GBM in a normal human glomerulus. E and F: two representative glomeruli from different patients with FSGS showing agrin staining along Bowman’s capsule. E′ and F′: higher magnification of the insets showing LAMB2 staining (white arrows) along Bowman’s capsule. G–I: COL4A4 (green) and DAPI. G: COL4A4 localized to the GBM in a normal human glomerulus. H and I: two representative glomeruli from different patients with FSGS showing COL4A4 staining along Bowman’s capsule. H′ and I′: higher magnification of the insets showing COL4A4 staining (white arrows) along Bowman’s capsule. These results show that podocyte-specific extracellular matrix proteins are expressed de novo by PECs in human FSGS. Original magnification: ×200. Scale bars = 100 μm.

Journal: American Journal of Physiology - Renal Physiology

Article Title: Differential expression of parietal epithelial cell and podocyte extracellular matrix proteins in focal segmental glomerulosclerosis and diabetic nephropathy

doi: 10.1152/ajprenal.00266.2019

Figure Lengend Snippet: The podocyte-specific extracellular matrix proteins laminin-β2 (LAMB2), agrin, and collagen type IV-α4 (COL4A4) increased in parietal epithelial cells (PECs) in human focal segmental glomerulosclerosis (FSGS). A–C: LAMB2 (green) and DAPI (blue). A: LAMB2 localized to the glomerular basement membrane (GBM) in the normal human kidney. B and C: two representative glomeruli from different patients with FSGS showing LAMB2 staining along Bowman’s capsule. B′ and C′: higher magnification of the insets showing LAMB2 staining (white arrows) along Bowman’s capsule. D–F: agrin (green) and DAPI. D: agrin localized to the GBM in a normal human glomerulus. E and F: two representative glomeruli from different patients with FSGS showing agrin staining along Bowman’s capsule. E′ and F′: higher magnification of the insets showing LAMB2 staining (white arrows) along Bowman’s capsule. G–I: COL4A4 (green) and DAPI. G: COL4A4 localized to the GBM in a normal human glomerulus. H and I: two representative glomeruli from different patients with FSGS showing COL4A4 staining along Bowman’s capsule. H′ and I′: higher magnification of the insets showing COL4A4 staining (white arrows) along Bowman’s capsule. These results show that podocyte-specific extracellular matrix proteins are expressed de novo by PECs in human FSGS. Original magnification: ×200. Scale bars = 100 μm.

Techniques: Membrane, Staining

Differential immunofluorescent immunostaining in parietal epithelial cells (PECs) for laminin-β2 (LAMB2), agrin, and collagen type IV-α4 (COL4A4) in an experimental diabetic nephropathy model. A−C: nondiabetic wild-type (WT) mice. Immunofluorescent staining for LAMB2 (A), agrin (B), and COL4A4 (C) staining (green) showed a podocyte distribution. Nuclei were stained with DAPI (blue). D−F: diabetic ob/ob mice. De novo staining was detected in PECs (white arrows) for LAMB2 (D), agrin (E), and COL4A4 (F) and in the mesangium (yellow arrows). G−I: quantification. The percentage of glomerular cross sections with de novo expression along Bowman’s capsule (BC) increased in diabetic mice for LAMB2 (G) and agrin (H) but not COL4A4 (I). Original magnification: ×400. Scale bars = 20 μm.

Journal: American Journal of Physiology - Renal Physiology

Article Title: Differential expression of parietal epithelial cell and podocyte extracellular matrix proteins in focal segmental glomerulosclerosis and diabetic nephropathy

doi: 10.1152/ajprenal.00266.2019

Figure Lengend Snippet: Differential immunofluorescent immunostaining in parietal epithelial cells (PECs) for laminin-β2 (LAMB2), agrin, and collagen type IV-α4 (COL4A4) in an experimental diabetic nephropathy model. A−C: nondiabetic wild-type (WT) mice. Immunofluorescent staining for LAMB2 (A), agrin (B), and COL4A4 (C) staining (green) showed a podocyte distribution. Nuclei were stained with DAPI (blue). D−F: diabetic ob/ob mice. De novo staining was detected in PECs (white arrows) for LAMB2 (D), agrin (E), and COL4A4 (F) and in the mesangium (yellow arrows). G−I: quantification. The percentage of glomerular cross sections with de novo expression along Bowman’s capsule (BC) increased in diabetic mice for LAMB2 (G) and agrin (H) but not COL4A4 (I). Original magnification: ×400. Scale bars = 20 μm.

Techniques: Immunostaining, Staining, Expressing

Laminin-β2 (LAMB2), agrin, and collagen type IV-α4 (COL4A4) staining in human diabetic nephropathy. A–C: LAMB2 staining. A: LAMB2 staining was restricted to the glomerular basement membrane (GBM) in the normal glomerulus. B and C: LAMB2 was detected along Bowman’s capsule in glomeruli from two different patients with diabetic nephropathy. B′ and C′: higher magnification of the insets showing de novo staining for LAMB2 along Bowman’s capsule (white arrows) and in the mesangium (yellow arrow). D–F: agrin staining. D: agrin staining localized to the GBM in the normal glomerulus. E and F: agrin was also detected along Bowman’s capsule in glomeruli from two different patients. E′ and F′: higher magnification of the insets showing staining for agrin along Bowman’s capsule (white arrows) and in the mesangium (yellow arrow). G–I: COL4A4 staining. G: COL4A4 staining localized to the GBM in the normal glomerulus. H and I: COL4A4 was also detected along Bowman’s capsule in glomeruli from two different patients with diabetic nephropathy. H′ and I′: higher magnification showing COL4A4 staining along Bowman’s capsule (white arrows) and in the mesangium (yellow arrow). Original magnification: ×200. Scale bars = 100 μm.

Journal: American Journal of Physiology - Renal Physiology

Article Title: Differential expression of parietal epithelial cell and podocyte extracellular matrix proteins in focal segmental glomerulosclerosis and diabetic nephropathy

doi: 10.1152/ajprenal.00266.2019

Figure Lengend Snippet: Laminin-β2 (LAMB2), agrin, and collagen type IV-α4 (COL4A4) staining in human diabetic nephropathy. A–C: LAMB2 staining. A: LAMB2 staining was restricted to the glomerular basement membrane (GBM) in the normal glomerulus. B and C: LAMB2 was detected along Bowman’s capsule in glomeruli from two different patients with diabetic nephropathy. B′ and C′: higher magnification of the insets showing de novo staining for LAMB2 along Bowman’s capsule (white arrows) and in the mesangium (yellow arrow). D–F: agrin staining. D: agrin staining localized to the GBM in the normal glomerulus. E and F: agrin was also detected along Bowman’s capsule in glomeruli from two different patients. E′ and F′: higher magnification of the insets showing staining for agrin along Bowman’s capsule (white arrows) and in the mesangium (yellow arrow). G–I: COL4A4 staining. G: COL4A4 staining localized to the GBM in the normal glomerulus. H and I: COL4A4 was also detected along Bowman’s capsule in glomeruli from two different patients with diabetic nephropathy. H′ and I′: higher magnification showing COL4A4 staining along Bowman’s capsule (white arrows) and in the mesangium (yellow arrow). Original magnification: ×200. Scale bars = 100 μm.

Techniques: Staining, Membrane

Podocyte-specific proteins colocalize with the activated parietal epithelial cell (PEC) marker CD44 in mice with experimental focal segmental glomerulosclerosis (FSGS). Double staining was performed for CD44 (red) and podocyte-specific extracellular matrix proteins (green). A−C: normal mice. The merged images show staining for laminin-β2 (LAMB2; A), agrin (B), and collagen type IV-α4 (COL4A4; C) but not CD44 in normal glomeruli. A1−C3: individual stains for LAMB2 (A1−A3), agrin (B1−B3), and COL4A4 (C1–C3). D−F: FSGS mice. The merged images show colocalization (yellow) of CD44 in PECs with LAMB2 (D and D3) and agrin (E and E3) but not COL4A4 (F and F3). D1, D2, E1, E2, F1, and F2: corresponding single stainings for LAMB2 (D1 and D2), agrin (E1 and E2), and COL4A4 (F1 and F2). Single and merged higher magnifications of the insets are shown in D4–F6). These results are consistent with activated PECs expressing the podocyte-specific extracellular matrix proteins LAMB2 and agrin in FSGS. Original magnification: ×400. Scale bars = 20 μm.

Journal: American Journal of Physiology - Renal Physiology

Article Title: Differential expression of parietal epithelial cell and podocyte extracellular matrix proteins in focal segmental glomerulosclerosis and diabetic nephropathy

doi: 10.1152/ajprenal.00266.2019

Figure Lengend Snippet: Podocyte-specific proteins colocalize with the activated parietal epithelial cell (PEC) marker CD44 in mice with experimental focal segmental glomerulosclerosis (FSGS). Double staining was performed for CD44 (red) and podocyte-specific extracellular matrix proteins (green). A−C: normal mice. The merged images show staining for laminin-β2 (LAMB2; A), agrin (B), and collagen type IV-α4 (COL4A4; C) but not CD44 in normal glomeruli. A1−C3: individual stains for LAMB2 (A1−A3), agrin (B1−B3), and COL4A4 (C1–C3). D−F: FSGS mice. The merged images show colocalization (yellow) of CD44 in PECs with LAMB2 (D and D3) and agrin (E and E3) but not COL4A4 (F and F3). D1, D2, E1, E2, F1, and F2: corresponding single stainings for LAMB2 (D1 and D2), agrin (E1 and E2), and COL4A4 (F1 and F2). Single and merged higher magnifications of the insets are shown in D4–F6). These results are consistent with activated PECs expressing the podocyte-specific extracellular matrix proteins LAMB2 and agrin in FSGS. Original magnification: ×400. Scale bars = 20 μm.

Techniques: Marker, Double Staining, Staining, Expressing

Autophagy promotes tubule-like structure formation by GSCs in 3D scaffold. (A) GSCs transfected with EGFP-LC3 were cultured in DMEM, STM or EBM for 72 h. Confocal microscopy was used to assess autophagy shown as green spots (left panel). Two hundred cells were randomly chosen to count the number of EGFP-LC3 fluorescent spots (right panel). Scale bar: 10 μm. (B) Confocal microscopy showing EGFP-transfected GSCs to form tubule-like structure in 3D scaffold (left panel). The number of tubular structures formed by GSCs in 3D scaffold was analyzed (right pannel). Scale bar: 20 μm. Asterisks (*) indicate VM channel formed by tumor cells. (C) Expression of autophagy proteins, ATG5, ATG7, LC3-I to LC3-II conversion and SQSTM1. TUBB/β-Tubulin served as a control. (D) The expressions of PROM1, GFAP, KDR proteins and the phosphorylation of Y1175 in the C-terminal region of KDR (p-KDR) as well as CDH5 and LAMB2. (E) Double staining of LC3 and CDH5 as well as LC3 and LAMB2 in GSCs cultured with EBM on the scaffold. LC3 is marked as red spots in the cytoplasm; CDH5 and LAMB2 are in green color in the cytoplasm. Scale bar: 20 μm. Nuclei were stained by Hoechst 33324 in blue. The results are expressed as the mean ± SEM from 3 experiments.

Journal: Autophagy

Article Title: Autophagy-induced KDR/VEGFR-2 activation promotes the formation of vasculogenic mimicry by glioma stem cells

doi: 10.1080/15548627.2017.1336277

Figure Lengend Snippet: Autophagy promotes tubule-like structure formation by GSCs in 3D scaffold. (A) GSCs transfected with EGFP-LC3 were cultured in DMEM, STM or EBM for 72 h. Confocal microscopy was used to assess autophagy shown as green spots (left panel). Two hundred cells were randomly chosen to count the number of EGFP-LC3 fluorescent spots (right panel). Scale bar: 10 μm. (B) Confocal microscopy showing EGFP-transfected GSCs to form tubule-like structure in 3D scaffold (left panel). The number of tubular structures formed by GSCs in 3D scaffold was analyzed (right pannel). Scale bar: 20 μm. Asterisks (*) indicate VM channel formed by tumor cells. (C) Expression of autophagy proteins, ATG5, ATG7, LC3-I to LC3-II conversion and SQSTM1. TUBB/β-Tubulin served as a control. (D) The expressions of PROM1, GFAP, KDR proteins and the phosphorylation of Y1175 in the C-terminal region of KDR (p-KDR) as well as CDH5 and LAMB2. (E) Double staining of LC3 and CDH5 as well as LC3 and LAMB2 in GSCs cultured with EBM on the scaffold. LC3 is marked as red spots in the cytoplasm; CDH5 and LAMB2 are in green color in the cytoplasm. Scale bar: 20 μm. Nuclei were stained by Hoechst 33324 in blue. The results are expressed as the mean ± SEM from 3 experiments.

Article Snippet: Antibodies Antibodies used include mouse anti-LAMB2/laminin B2 monoclonal antibody (mAb; BD Biosciences, 610722), rabbit anti-CDH5/cadherin 5 mAb (Cell Signaling Technology, 2158), rabbit anti-PROM1/CD133 polyclonal antibody (pAb; Santa Cruz Biotechnology, sc-30220), rabbit anti-GFAP pAb (Abcam, ab7260), mouse anti-CD34 mAb (Abcam, ab187282), rabbit anti-VEGF pAb (Proteintech, 19003–1-AP), rabbit anti-phospho- KDR/VEGFR-2 mAb (Cell Signaling Technology, 2478), rabbit anti-KDR/VEGFR-2 mAb (Cell Signaling Technology, 2479), mouse anti-LC3B mAb (Santa Cruz Biotechnology, sc-376404), rabbit anti-ATG5 mAb (Cell Signaling Technology, 8540), rabbit anti-ATG7 mAb (Cell Signaling Technology, 8558), rabbit anti-SQSTM1/p62 pAb (Sigma-Aldrich, P0067), rabbit anti-BECN1 mAb (Abcam, ab51031), mouse anti-PI3K p85 mAb (Abcam, ab86714), rabbit anti-pan-AKT pAb (Abcam, ab8805), rabbit anti-phospho-PI3K p85 (Y607) pAb (Abcam, ab182651), rabbit anti-phospho-pan-AKT pAb (Abcam, ab38449).

Techniques: Transfection, Cell Culture, Confocal Microscopy, Expressing, Double Staining, Staining

Tubular structure formation by GSCs is promoted by rapamycin (RAPA). (A) RAPA (100 nM) was added to EBM and confocal microscopy was used to detect autophagic vesicles in GSCs forming tubular structure (left panel). Two hundred cells were randomly chosen to count green fluorescent spots within the cells (right panel). Scale bar: 10 μm. (B) Confocal microscopy showing EGFP-transfected GSCs to form tubule-like structure in 3D scaffold (left panel). The number of tubular structures was counted in the scaffold cultured with or without RAPA (right panel). Scale bar: 20 μm. Asterisks (*) indicate VM channel formed by tumor cells. (C) The expression of LC3-I to LC3-II conversion, SQSTM1 and VM-related proteins (KDR, p-KDR, CDH5 and LAMB2) in GSCs treated by RAPA. The results are expressed as the mean ± SEM from 3 experiments.

Journal: Autophagy

Article Title: Autophagy-induced KDR/VEGFR-2 activation promotes the formation of vasculogenic mimicry by glioma stem cells

doi: 10.1080/15548627.2017.1336277

Figure Lengend Snippet: Tubular structure formation by GSCs is promoted by rapamycin (RAPA). (A) RAPA (100 nM) was added to EBM and confocal microscopy was used to detect autophagic vesicles in GSCs forming tubular structure (left panel). Two hundred cells were randomly chosen to count green fluorescent spots within the cells (right panel). Scale bar: 10 μm. (B) Confocal microscopy showing EGFP-transfected GSCs to form tubule-like structure in 3D scaffold (left panel). The number of tubular structures was counted in the scaffold cultured with or without RAPA (right panel). Scale bar: 20 μm. Asterisks (*) indicate VM channel formed by tumor cells. (C) The expression of LC3-I to LC3-II conversion, SQSTM1 and VM-related proteins (KDR, p-KDR, CDH5 and LAMB2) in GSCs treated by RAPA. The results are expressed as the mean ± SEM from 3 experiments.

Techniques: Confocal Microscopy, Transfection, Cell Culture, Expressing

Autophagy mediates the formation of VM through KDR phosphorylation. (A) Confocal microscopy detection of autophagic vesicles formed by GSCs transfected with EGFP-LC3 and treated with 5 μM chloroquine (CQ) (left panel). Two hundred cells were randomly chosen to count green fluorescent spots within the cells (right panel). Scale bar: 10 μm. (B) Confocal microscopy showing EGFP-transfected GSCs to form vascular tubules in 3D scaffold (left panel). The number of tubular structures was counted in the scaffold with or without CQ (right panel). Scale bar: 20 μm. Asterisks (*) indicate VM channels formed by tumor cells. (C) The expression of LC3-I to LC3-II conversion, SQSTM1 and KDR, p-KDR, LAMB2, and CDH5 in GSCs after 5 μM CQ treatment. (D) Western blot detection of p-KDR, KDR, LAMB2, CDH5 and SQSTM1 in GSCs in medium with 100 μg/ml BEV and/or 5 μM CQ. The results are expressed as the mean ± SEM from 3 experiments.

Journal: Autophagy

Article Title: Autophagy-induced KDR/VEGFR-2 activation promotes the formation of vasculogenic mimicry by glioma stem cells

doi: 10.1080/15548627.2017.1336277

Figure Lengend Snippet: Autophagy mediates the formation of VM through KDR phosphorylation. (A) Confocal microscopy detection of autophagic vesicles formed by GSCs transfected with EGFP-LC3 and treated with 5 μM chloroquine (CQ) (left panel). Two hundred cells were randomly chosen to count green fluorescent spots within the cells (right panel). Scale bar: 10 μm. (B) Confocal microscopy showing EGFP-transfected GSCs to form vascular tubules in 3D scaffold (left panel). The number of tubular structures was counted in the scaffold with or without CQ (right panel). Scale bar: 20 μm. Asterisks (*) indicate VM channels formed by tumor cells. (C) The expression of LC3-I to LC3-II conversion, SQSTM1 and KDR, p-KDR, LAMB2, and CDH5 in GSCs after 5 μM CQ treatment. (D) Western blot detection of p-KDR, KDR, LAMB2, CDH5 and SQSTM1 in GSCs in medium with 100 μg/ml BEV and/or 5 μM CQ. The results are expressed as the mean ± SEM from 3 experiments.

Techniques: Confocal Microscopy, Transfection, Expressing, Western Blot

Knockdown of ATG5 reduces tubular structure formation by GSCs and phosphorylation of KDR. (A) The expression of ATG5, p-KDR, KDR, CDH5 and LAMB2 in GSCs transfected with siATG5. (B) Two hundred cells were randomly chosen to count green fluorescent spots within the cells. (C) The number of tubular structures formed by GSCs and siATG5-GSCs in the scaffold counted with confocal microscopy. (D) Western blot detection of p-KDR and KDR in siATG5-GSCs cultured with 100 μg/ml BEV. (E) and (F) Mice with intracranial injection of GSCs transfected with siCtrl RNA or siATG5 were treated with BEV. Forty-two animals were then randomly divided into 7 groups consisting of 6 animals each. After 4 wk, tumors were collected to analyze vessel density (E) and formation of VM (F). HPF, high-power field. The results are expressed as the mean ± SEM from 3 experiments.

Journal: Autophagy

Article Title: Autophagy-induced KDR/VEGFR-2 activation promotes the formation of vasculogenic mimicry by glioma stem cells

doi: 10.1080/15548627.2017.1336277

Figure Lengend Snippet: Knockdown of ATG5 reduces tubular structure formation by GSCs and phosphorylation of KDR. (A) The expression of ATG5, p-KDR, KDR, CDH5 and LAMB2 in GSCs transfected with siATG5. (B) Two hundred cells were randomly chosen to count green fluorescent spots within the cells. (C) The number of tubular structures formed by GSCs and siATG5-GSCs in the scaffold counted with confocal microscopy. (D) Western blot detection of p-KDR and KDR in siATG5-GSCs cultured with 100 μg/ml BEV. (E) and (F) Mice with intracranial injection of GSCs transfected with siCtrl RNA or siATG5 were treated with BEV. Forty-two animals were then randomly divided into 7 groups consisting of 6 animals each. After 4 wk, tumors were collected to analyze vessel density (E) and formation of VM (F). HPF, high-power field. The results are expressed as the mean ± SEM from 3 experiments.

Techniques: Expressing, Transfection, Confocal Microscopy, Western Blot, Cell Culture, Injection

Review

Structured Review

Images

Similar Products

Image Search Results