Journal: STAR Protocols

Article Title: Simultaneous isolation of CD45 tumor-infiltrating lymphocytes, tumor cells, and associated fibroblasts from murine breast tumor model by MACS

doi: 10.1016/j.xpro.2022.101951

Figure Lengend Snippet:

Article Snippet: CD140b Antibody, anti-mouse, REAfinity- APC-770 (1:50 dilution) , Miltenyi Biotec , Cat# 130-118-469, RRID: AB_2733095.

Techniques: Recombinant, Flow Cytometry, Staining, Isolation, Cell Isolation, Software, Real-time Polymerase Chain Reaction, Blocking Assay, Gentle

Journal: Molecular medicine reports

Article Title: Protective effects of glutamine in a rat model of endotoxemia.

doi: 10.3892/mmr.2012.1007

Figure Lengend Snippet: Figure 4. Immunolocalization of (A-C) platelet-derived growth factor-B (PDGF-B) and (D-F) PDGF receptor-β (PDGFR-β) in brain tissue of (A,D) controls, (B,E) lipopolysaccharide (LPS) group at 72 h, and the (C,F) glutamine (Gln) treatment group at 72 h. PDGF-B immunoreactivity was observed in the cyto plasm, PDGFR-β immunoreactivity was observed in the membrane of cortical neurons.

Article Snippet: Rabbit anti-rat NF-κB, PDGF-B, and PDGFR-β antibodies and ABC kits were purchased from Boster (Wuhan, China).

Techniques: Derivative Assay, Membrane

Journal: Molecular medicine reports

Article Title: Protective effects of glutamine in a rat model of endotoxemia.

doi: 10.3892/mmr.2012.1007

Figure Lengend Snippet: Figure 6. Western blot analysis of PDGF receptor-β (PDGFR-β) protein at various time points following injection of lipopolysaccharide (LPS). Upper row depicts the control group from left to right as 1-5 (2, 6, 12, 24 and 72 h) and the treatment group as 6-10 (2, 6, 12, 24 and 72 h). Lower row depicts the control group from left to right as 1-5 (2, 6, 12, 24 and 72 h) and the LPS group as 6-10 (2, 6, 12, 24 and 72 h).

Article Snippet: Rabbit anti-rat NF-κB, PDGF-B, and PDGFR-β antibodies and ABC kits were purchased from Boster (Wuhan, China).

Techniques: Western Blot, Injection, Control

Journal: Cell Death & Disease

Article Title: Intrinsic regulation of hemangioma involution by platelet-derived growth factor

doi: 10.1038/cddis.2012.58

Figure Lengend Snippet: Expression of PDGF receptors in proliferating and involuting hemangiomas. ( a ) Real-time RT-PCR analysis of PDGFR- α , - β , and PPAR γ in proliferating and involuting hemangiomas (data normalized to 18S rRNA and presented as relative to normal skin; * P <0.05 compared with normal skin, † P <0.05 compared with proliferating hemangioma; n =3). ( b ) Immunostaining of proliferating hemangiomas for PDGFR- α and PDGFR- β (images taken at × 20, inserts show higher magnification; brown=DAB staining, blue=hematoxylin). ( c ) Immunofluorescence double labeling for mesenchymal cell marker α -SMA and PDGFR- β (images taken at × 20, green=PDGFR- β , red= α -SMA)

Article Snippet: The cells were transfected with control shRNA (sc-108060; Santa Cruz Biotechnology), PDGFR- α shRNA (sc-29443-SH; Santa Cruz Biotechnology) plasmid DNA, or PDGFR- β shRNA (sc-29442-SH; Santa Cruz Biotechnology) at a concentration of 0.25 μ g/cm 2 surface area.

Techniques: Expressing, Quantitative RT-PCR, Immunostaining, Staining, Immunofluorescence, Labeling, Marker

Journal: Cell Death & Disease

Article Title: Intrinsic regulation of hemangioma involution by platelet-derived growth factor

doi: 10.1038/cddis.2012.58

Figure Lengend Snippet: Phosphorylated PDGF receptors in proliferating hemangiomas. Immunostaining of proliferating hemangioma specimens for phospho-PDGFR- β ( a ) and phospho-PDGFR- α ( b ) (images taken at × 20, green=PDGFR, blue=DAPI)

Article Snippet: The cells were transfected with control shRNA (sc-108060; Santa Cruz Biotechnology), PDGFR- α shRNA (sc-29443-SH; Santa Cruz Biotechnology) plasmid DNA, or PDGFR- β shRNA (sc-29442-SH; Santa Cruz Biotechnology) at a concentration of 0.25 μ g/cm 2 surface area.

Techniques: Immunostaining

Journal: Cell Death & Disease

Article Title: Intrinsic regulation of hemangioma involution by platelet-derived growth factor

doi: 10.1038/cddis.2012.58

Figure Lengend Snippet: Expression of PDGF signaling axis in hemangioma-derived CD133+ cells. ( a ) Expression of PDGFRs in hemSCs and bm-MPCs (data normalized to 18 S rRNA and presented as relative to bm-MPCs; two different hemSC preparations run in triplicates were averaged). ( b ) Expression of PDGF transcripts in hemSCs (data presented as in ( a ); * P <0.05 compared with bm-MPCs; right panel shows the expression of PDGF-A, PDGF-C, and PDGF-D at a lower scale). ( c ) PDGF-BB levels in media from cells cultured for 48 h in EBM2/20%FBS (with growth factors) (* P <0.05 compared with bm-MPCs). ( d ) PDGF-BB levels in cell lysates determined by ELISA (* P <0.05 compared with bm-MPCs). ( e ) Levels of phosphorylated PDGFR- α and - β in hemSCs with or without exogenous PDGF-BB stimulation (cells were stimulated with 10 ng/ml PDGF-BB; * P <0.05 compared with unstimulated cells). ( f ) Schematic illustration of the hypothesized PDGF polypeptides and the corresponding receptors on hemangioma stem cell surface

Article Snippet: The cells were transfected with control shRNA (sc-108060; Santa Cruz Biotechnology), PDGFR- α shRNA (sc-29443-SH; Santa Cruz Biotechnology) plasmid DNA, or PDGFR- β shRNA (sc-29442-SH; Santa Cruz Biotechnology) at a concentration of 0.25 μ g/cm 2 surface area.

Techniques: Expressing, Derivative Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Cell Death & Disease

Article Title: Intrinsic regulation of hemangioma involution by platelet-derived growth factor

doi: 10.1038/cddis.2012.58

Figure Lengend Snippet: PDGF-BB inhibits adipogenic differentiation in hemSCs. ( a ) Representative oil red O staining of hemSCs cultured in adipogenic differentiation media supplemented with 10 ng/ml of PDGF-AA, -AB, or -BB. ( b ) Quantitative analysis of adipogenic differentiation in the presence of PDGF polypeptides was assessed by real-time RT-PCR for adipogenesis-specific transcription factors, C/EBP α and PPAR γ (data normalized to 18 S rRNA and presented as relative to adipogenic media; * P <0.05 compared with the adipogenic media; n =3). ( c ) Effect of adipogenic differentiation on PDGFR expression in hemSCs at day 7. ( d ) Oil red O staining of hemSCs cultured with different growth factors (each at 10 ng/ml) illustrating specific inhibition of hemSC adipogenesis by PDGF signaling

Article Snippet: The cells were transfected with control shRNA (sc-108060; Santa Cruz Biotechnology), PDGFR- α shRNA (sc-29443-SH; Santa Cruz Biotechnology) plasmid DNA, or PDGFR- β shRNA (sc-29442-SH; Santa Cruz Biotechnology) at a concentration of 0.25 μ g/cm 2 surface area.

Techniques: Staining, Cell Culture, Quantitative RT-PCR, Expressing, Inhibition

Journal: Cell Death & Disease

Article Title: Intrinsic regulation of hemangioma involution by platelet-derived growth factor

doi: 10.1038/cddis.2012.58

Figure Lengend Snippet: Inhibiting cell-autogenous PDGF signaling enhances adipogenesis. ( a ) Oil red O staining of hemSCs exposed to adipogenic media with or without PDGFR inhibitors, AG-370 and AG-1296, and PDGFR neutralizing antibodies. ( b ) Quantitative analysis of C/EBP α and PPAR γ in cells exposed to PDGF inhibitors as in ( a ) (data presented relative to control media; * P <0.05 compared with control media; † P <0.05 compared with adipogenic media; n =3). ( c ) Effect of PDGFR- α and PDGFR- β- neutralizing antibodies and chemical inhibitors on reversing the inhibition by exogenous PDGF-BB (* P <0.05 compared with PDGF-BB treatment)

Article Snippet: The cells were transfected with control shRNA (sc-108060; Santa Cruz Biotechnology), PDGFR- α shRNA (sc-29443-SH; Santa Cruz Biotechnology) plasmid DNA, or PDGFR- β shRNA (sc-29442-SH; Santa Cruz Biotechnology) at a concentration of 0.25 μ g/cm 2 surface area.

Techniques: Staining, Control, Inhibition

Journal: Cell Death & Disease

Article Title: Intrinsic regulation of hemangioma involution by platelet-derived growth factor

doi: 10.1038/cddis.2012.58

Figure Lengend Snippet: PDGF-BB employs PDGFR- β in hemSCs. shRNA-mediated knockdown of PDGFR- α ( a ) and PDGFR- β ( b ) in hemSCs at sub-passage 1 (subP1; following 2 weeks of puromycin selection) and sub-passage 6 (six serial passages after puromycin selection) (* P <0.05 compared with control shRNA). ( c ) shRNA-knockdown of PDGFR- α and PDGFR- β in hemSCs reduced the inhibitory effect of PDGF-BB on hemSC adipogenesis (* P <0.05 compared with control shRNA). ( d ) Schematic illustrating the findings of the study and our working hypothesis. High levels of PDGF-BB from hemSCs during proliferation mediate intracellular signaling through PDGFR- β (predominantly PDGFR- ββ homodimer with contribution from PDGFR- αβ heterodimer) to inhibit C/EBP α and PPAR γ expression and adipogenesis (i.e., involution). Upon regression of the vessels and removal of the PDGF-BB source, adipogenesis is triggered

Article Snippet: The cells were transfected with control shRNA (sc-108060; Santa Cruz Biotechnology), PDGFR- α shRNA (sc-29443-SH; Santa Cruz Biotechnology) plasmid DNA, or PDGFR- β shRNA (sc-29442-SH; Santa Cruz Biotechnology) at a concentration of 0.25 μ g/cm 2 surface area.

Techniques: shRNA, Knockdown, Selection, Control, Expressing