hif1a Search Results


gene exp hif1a mm00468869 m1  (Thermo Fisher)
99
Thermo Fisher gene exp hif1a mm00468869 m1
Gene Exp Hif1a Mm00468869 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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gene exp hif1a bt03259341 m1  (Thermo Fisher)
94
Thermo Fisher gene exp hif1a bt03259341 m1
Gene Exp Hif1a Bt03259341 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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hif1α monoclonal antibody  (Boster Bio)
90
Boster Bio hif1α monoclonal antibody
Hif1α Monoclonal Antibody, supplied by Boster Bio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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antibody anti hif1a  (R&D Systems)
95
R&D Systems antibody anti hif1a
Figure 6. Rescue of Tsc1fl/fl;Tyrp1-Cre mouse retinal phenotypes by concomitant deletion of hypoxia-induced factor 1-alpha <t>(Hif1a).</t> (A) Levels of senescence markers in P30 mouse retinas with indicated genotypes were analyzed by Western blot (WB). Relative amounts of proteins used in each sample were determined by WB detection of β-actin. (B) Distributions of mTORC1-active cells, which are positive to pS6, MG, which are positive to Sox9, and senescent cells, which are positive to β-gal and p53, were examined by immunostaining of the eye sections. Expression of those markers in the Cre-affected cells were determined by comparing the expression of the Cre reporter R26tdTom. (C) Numbers of Sox9-positive MG in the mouse retinas were counted and the relative numbers are shown in the graph. (D) R26tdTom-negative wild-type cells and R26tdTom-positive Cre-affected cells in the retinas were determined byFACS and shown in the graph. (E) The lengths of telomeres of the cells isolated from P30 mouse retinas with the indicated genotypes were compared with that of control sample, and their relative values are shown in the graph. Error bars denote SD. Numbers of samples analyzed are shown in the graphs. n.s., not significant; *p < 0.05; **p < 0.01; ***p < 0.001.
Antibody Anti Hif1a, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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short hairpin rna shrna constructs against mus musculus hif 1a  (OriGene)
90
OriGene short hairpin rna shrna constructs against mus musculus hif 1a
Figure 6. Rescue of Tsc1fl/fl;Tyrp1-Cre mouse retinal phenotypes by concomitant deletion of hypoxia-induced factor 1-alpha <t>(Hif1a).</t> (A) Levels of senescence markers in P30 mouse retinas with indicated genotypes were analyzed by Western blot (WB). Relative amounts of proteins used in each sample were determined by WB detection of β-actin. (B) Distributions of mTORC1-active cells, which are positive to pS6, MG, which are positive to Sox9, and senescent cells, which are positive to β-gal and p53, were examined by immunostaining of the eye sections. Expression of those markers in the Cre-affected cells were determined by comparing the expression of the Cre reporter R26tdTom. (C) Numbers of Sox9-positive MG in the mouse retinas were counted and the relative numbers are shown in the graph. (D) R26tdTom-negative wild-type cells and R26tdTom-positive Cre-affected cells in the retinas were determined byFACS and shown in the graph. (E) The lengths of telomeres of the cells isolated from P30 mouse retinas with the indicated genotypes were compared with that of control sample, and their relative values are shown in the graph. Error bars denote SD. Numbers of samples analyzed are shown in the graphs. n.s., not significant; *p < 0.05; **p < 0.01; ***p < 0.001.
Short Hairpin Rna Shrna Constructs Against Mus Musculus Hif 1a, supplied by OriGene, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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hif 1α sirna  (OriGene)
90
OriGene hif 1α sirna
Figure 6. Rescue of Tsc1fl/fl;Tyrp1-Cre mouse retinal phenotypes by concomitant deletion of hypoxia-induced factor 1-alpha <t>(Hif1a).</t> (A) Levels of senescence markers in P30 mouse retinas with indicated genotypes were analyzed by Western blot (WB). Relative amounts of proteins used in each sample were determined by WB detection of β-actin. (B) Distributions of mTORC1-active cells, which are positive to pS6, MG, which are positive to Sox9, and senescent cells, which are positive to β-gal and p53, were examined by immunostaining of the eye sections. Expression of those markers in the Cre-affected cells were determined by comparing the expression of the Cre reporter R26tdTom. (C) Numbers of Sox9-positive MG in the mouse retinas were counted and the relative numbers are shown in the graph. (D) R26tdTom-negative wild-type cells and R26tdTom-positive Cre-affected cells in the retinas were determined byFACS and shown in the graph. (E) The lengths of telomeres of the cells isolated from P30 mouse retinas with the indicated genotypes were compared with that of control sample, and their relative values are shown in the graph. Error bars denote SD. Numbers of samples analyzed are shown in the graphs. n.s., not significant; *p < 0.05; **p < 0.01; ***p < 0.001.
Hif 1α Sirna, supplied by OriGene, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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hif 1α mab1536  (R&D Systems)
95
R&D Systems hif 1α mab1536
Figure 6. Rescue of Tsc1fl/fl;Tyrp1-Cre mouse retinal phenotypes by concomitant deletion of hypoxia-induced factor 1-alpha <t>(Hif1a).</t> (A) Levels of senescence markers in P30 mouse retinas with indicated genotypes were analyzed by Western blot (WB). Relative amounts of proteins used in each sample were determined by WB detection of β-actin. (B) Distributions of mTORC1-active cells, which are positive to pS6, MG, which are positive to Sox9, and senescent cells, which are positive to β-gal and p53, were examined by immunostaining of the eye sections. Expression of those markers in the Cre-affected cells were determined by comparing the expression of the Cre reporter R26tdTom. (C) Numbers of Sox9-positive MG in the mouse retinas were counted and the relative numbers are shown in the graph. (D) R26tdTom-negative wild-type cells and R26tdTom-positive Cre-affected cells in the retinas were determined byFACS and shown in the graph. (E) The lengths of telomeres of the cells isolated from P30 mouse retinas with the indicated genotypes were compared with that of control sample, and their relative values are shown in the graph. Error bars denote SD. Numbers of samples analyzed are shown in the graphs. n.s., not significant; *p < 0.05; **p < 0.01; ***p < 0.001.
Hif 1α Mab1536, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti hif 1α  (R&D Systems)
93
R&D Systems anti hif 1α
Figure 6. Rescue of Tsc1fl/fl;Tyrp1-Cre mouse retinal phenotypes by concomitant deletion of hypoxia-induced factor 1-alpha <t>(Hif1a).</t> (A) Levels of senescence markers in P30 mouse retinas with indicated genotypes were analyzed by Western blot (WB). Relative amounts of proteins used in each sample were determined by WB detection of β-actin. (B) Distributions of mTORC1-active cells, which are positive to pS6, MG, which are positive to Sox9, and senescent cells, which are positive to β-gal and p53, were examined by immunostaining of the eye sections. Expression of those markers in the Cre-affected cells were determined by comparing the expression of the Cre reporter R26tdTom. (C) Numbers of Sox9-positive MG in the mouse retinas were counted and the relative numbers are shown in the graph. (D) R26tdTom-negative wild-type cells and R26tdTom-positive Cre-affected cells in the retinas were determined byFACS and shown in the graph. (E) The lengths of telomeres of the cells isolated from P30 mouse retinas with the indicated genotypes were compared with that of control sample, and their relative values are shown in the graph. Error bars denote SD. Numbers of samples analyzed are shown in the graphs. n.s., not significant; *p < 0.05; **p < 0.01; ***p < 0.001.
Anti Hif 1α, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ha hif1  (Addgene inc)
90
Addgene inc ha hif1
Figure 6. Rescue of Tsc1fl/fl;Tyrp1-Cre mouse retinal phenotypes by concomitant deletion of hypoxia-induced factor 1-alpha <t>(Hif1a).</t> (A) Levels of senescence markers in P30 mouse retinas with indicated genotypes were analyzed by Western blot (WB). Relative amounts of proteins used in each sample were determined by WB detection of β-actin. (B) Distributions of mTORC1-active cells, which are positive to pS6, MG, which are positive to Sox9, and senescent cells, which are positive to β-gal and p53, were examined by immunostaining of the eye sections. Expression of those markers in the Cre-affected cells were determined by comparing the expression of the Cre reporter R26tdTom. (C) Numbers of Sox9-positive MG in the mouse retinas were counted and the relative numbers are shown in the graph. (D) R26tdTom-negative wild-type cells and R26tdTom-positive Cre-affected cells in the retinas were determined byFACS and shown in the graph. (E) The lengths of telomeres of the cells isolated from P30 mouse retinas with the indicated genotypes were compared with that of control sample, and their relative values are shown in the graph. Error bars denote SD. Numbers of samples analyzed are shown in the graphs. n.s., not significant; *p < 0.05; **p < 0.01; ***p < 0.001.
Ha Hif1, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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immunofluorescence  (Proteintech)
96
Proteintech immunofluorescence
Figure 6. Rescue of Tsc1fl/fl;Tyrp1-Cre mouse retinal phenotypes by concomitant deletion of hypoxia-induced factor 1-alpha <t>(Hif1a).</t> (A) Levels of senescence markers in P30 mouse retinas with indicated genotypes were analyzed by Western blot (WB). Relative amounts of proteins used in each sample were determined by WB detection of β-actin. (B) Distributions of mTORC1-active cells, which are positive to pS6, MG, which are positive to Sox9, and senescent cells, which are positive to β-gal and p53, were examined by immunostaining of the eye sections. Expression of those markers in the Cre-affected cells were determined by comparing the expression of the Cre reporter R26tdTom. (C) Numbers of Sox9-positive MG in the mouse retinas were counted and the relative numbers are shown in the graph. (D) R26tdTom-negative wild-type cells and R26tdTom-positive Cre-affected cells in the retinas were determined byFACS and shown in the graph. (E) The lengths of telomeres of the cells isolated from P30 mouse retinas with the indicated genotypes were compared with that of control sample, and their relative values are shown in the graph. Error bars denote SD. Numbers of samples analyzed are shown in the graphs. n.s., not significant; *p < 0.05; **p < 0.01; ***p < 0.001.
Immunofluorescence, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/hif1a/HIF1a+Polyclonal+antibody/pmc12957808-96-5-10
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hif1α cat  (Proteintech)
96
Proteintech hif1α cat
Fig. 4. Signal pathway identification and transcription factor activity verification. A: Representative western blotting strips of PAs from MCT-induced PAH rats with or without metformin treatment and densitometric analysis showing <t>Hif1α,</t> PKG relative to β-actin; B: Representative western blotting strips of PAs from MCT- induced PAH rats with or without metformin treatment and densitometric analysis showing p-AKT relative to AKT; C: Representative western blotting strips of PAs from MCT-induced PAH rats with or without metformin treatment and densitometric analysis showing p53 relative to β-actin; D: Representative western blotting strips of PAs from MCT-induced PAH rats with or without metformin treatment and densitometric analysis showing Sp1, Est1 relative to β-actin. MCT means monocrotaline; Met means metformin. **p < 0.01. n = 3.
Hif1α Cat, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit polyclonal anti human mouse hif1α antibody  (ABclonal Biotechnology)
95
ABclonal Biotechnology rabbit polyclonal anti human mouse hif1α antibody
Fig. 4. Signal pathway identification and transcription factor activity verification. A: Representative western blotting strips of PAs from MCT-induced PAH rats with or without metformin treatment and densitometric analysis showing <t>Hif1α,</t> PKG relative to β-actin; B: Representative western blotting strips of PAs from MCT- induced PAH rats with or without metformin treatment and densitometric analysis showing p-AKT relative to AKT; C: Representative western blotting strips of PAs from MCT-induced PAH rats with or without metformin treatment and densitometric analysis showing p53 relative to β-actin; D: Representative western blotting strips of PAs from MCT-induced PAH rats with or without metformin treatment and densitometric analysis showing Sp1, Est1 relative to β-actin. MCT means monocrotaline; Met means metformin. **p < 0.01. n = 3.
Rabbit Polyclonal Anti Human Mouse Hif1α Antibody, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Figure 6. Rescue of Tsc1fl/fl;Tyrp1-Cre mouse retinal phenotypes by concomitant deletion of hypoxia-induced factor 1-alpha (Hif1a). (A) Levels of senescence markers in P30 mouse retinas with indicated genotypes were analyzed by Western blot (WB). Relative amounts of proteins used in each sample were determined by WB detection of β-actin. (B) Distributions of mTORC1-active cells, which are positive to pS6, MG, which are positive to Sox9, and senescent cells, which are positive to β-gal and p53, were examined by immunostaining of the eye sections. Expression of those markers in the Cre-affected cells were determined by comparing the expression of the Cre reporter R26tdTom. (C) Numbers of Sox9-positive MG in the mouse retinas were counted and the relative numbers are shown in the graph. (D) R26tdTom-negative wild-type cells and R26tdTom-positive Cre-affected cells in the retinas were determined byFACS and shown in the graph. (E) The lengths of telomeres of the cells isolated from P30 mouse retinas with the indicated genotypes were compared with that of control sample, and their relative values are shown in the graph. Error bars denote SD. Numbers of samples analyzed are shown in the graphs. n.s., not significant; *p < 0.05; **p < 0.01; ***p < 0.001.

Journal: eLife

Article Title: mTORC1-induced retinal progenitor cell overproliferation leads to accelerated mitotic aging and degeneration of descendent Müller glia

doi: 10.7554/elife.70079

Figure Lengend Snippet: Figure 6. Rescue of Tsc1fl/fl;Tyrp1-Cre mouse retinal phenotypes by concomitant deletion of hypoxia-induced factor 1-alpha (Hif1a). (A) Levels of senescence markers in P30 mouse retinas with indicated genotypes were analyzed by Western blot (WB). Relative amounts of proteins used in each sample were determined by WB detection of β-actin. (B) Distributions of mTORC1-active cells, which are positive to pS6, MG, which are positive to Sox9, and senescent cells, which are positive to β-gal and p53, were examined by immunostaining of the eye sections. Expression of those markers in the Cre-affected cells were determined by comparing the expression of the Cre reporter R26tdTom. (C) Numbers of Sox9-positive MG in the mouse retinas were counted and the relative numbers are shown in the graph. (D) R26tdTom-negative wild-type cells and R26tdTom-positive Cre-affected cells in the retinas were determined byFACS and shown in the graph. (E) The lengths of telomeres of the cells isolated from P30 mouse retinas with the indicated genotypes were compared with that of control sample, and their relative values are shown in the graph. Error bars denote SD. Numbers of samples analyzed are shown in the graphs. n.s., not significant; *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: 70079 18 of 27 Reagent type (species) or resource Designation Source or reference Identifiers Additional information Antibody Anti- BrdU(CldU) (Rat monoclonal) Novus Biologicals NB500- 169 IHC (1:200) Antibody Anti- BrdU(IdU) (Mouse monoclonal) EXBIO 11–286 C100 IHC (1:200) Antibody Anti- Brn3b (Rabbit polyclonal) Santa Cruz Biotechnology sc- 31989 IHC (1:200) Antibody Anti- Calbindin (Rabbit polyclonal) Swant Inc. CB- 38 IHC (1:200) Antibody Anti- Cdo (Goat polyclonal) R&D Systems AF2429 IHC (1:200) Antibody Anti- c- myc (Mouse monoclonal) Santa Cruz Biotechnology sc40 WB (1:1000) Antibody Anti- Cleaved caspase- 3 (Rabbit polyclonal) Cell Signaling Technology 9661 IHC (1:200) Antibody Anti- Ezrin (Mouse monoclonal) Invitrogen Biotechnology 35–7300 IHC (1:200) Antibody Anti- GFAP (Rabbit polyclonal) Abcam ab48050 IHC (1:200) Antibody Anti- GFP (Chicken polyclonal) Abcam ab13970 IHC (1:2000) Antibody Anti- Glutamine synthase (Rabbit polyclonal) Sigma- Aldrich G- 2781 IHC (1:200) Antibody Anti- Hif1a (Mouse monoclonal) R&D Systems MAB1536 WB (1:1000) Antibody Anti- Hif2α/Epas1 (Rabbit polyclonal) Novus Biologicals NB100- 122 WB (1:1000) Antibody Anti- Hk2 (Rabbit polyclonal) Cell Signaling Technology 2867 WB 1:1,000 IHC (1:200) Antibody Anti- Hydroxy- Hif1alpha (Rabbit polyclonal) Cell Signaling Technology 3434 WB (1:1000) Antibody Anti- M- opsin Merck Millipore AB5405 IHC (1:200) Antibody Anti- Otx2 (Rabbit polyclonal) Abcam ab25985 IHC (1:200) Antibody Anti- Otx2 (Rabbit polyclonal) Abcam ab183951 IHC (1:200) Antibody Anti- Otx2 (Goat polyclonal) R&D Systems AF1979- SP IHC (1:200) Antibody Anti- p21/Cip1 (Mouse monoclonal) Santa Cruz Biotechnology SC817 WB (1:1000) IHC (1:200) Antibody Anti- p53 (Mouse monoclonal) Merck Millipore OP03- 100 WB (1:1000) IHC (1:200) Antibody Anti- Pax6 (Rabbit polyclonal) Covance PRB- 278P IHC (1:200) Antibody Anti- Phospho Smad1/5(S463/465) (Rabbit polyclonal) Invitrogen Biotechnology 700047 IHC (1:200) Antibody Anti- phospho Smad2(ser465/467) (Rabbit polyclonal) - Cell Signaling Technology 18338 IHC (1:200) Antibody Anti- phospho- Histone H3(S10) (pH3; Rabbit polyclonal) Merck Millipore 04–1093 IHC (1:200) Antibody Anti- phospho- S6(S235/236) (pS6; Rabbit polyclonal) Cell Signaling Technology 2211 WB (1:1000) IHC (1:200) Continued Continued on next page Lim et al. eLife 2021;10:e70079.

Techniques: Western Blot, Immunostaining, Expressing, Isolation, Control

Figure 7. Hif1a supports mTORC1-induced retinal progenitor cell (RPC) proliferation through the expression of glycolytic enzymes. (A) The Cre-affected cells in P0 mouse retinas with the indicated genotypes were visualized by R26tdTom Cre reporter. Distribution of mTORC1-active cells in the boxed areas was also examined by immunostaining of pS6 and shown in the second row. Proliferation and cell cycle progression of RPCs in the mouse retinas were also examined by 5-ethynyl-2′-deoxyuridine (EdU)/5-chloro-2'-deoxyuridine (CldU) labeling and chasing experiments as described in Materials and methods. (B) R26tdTom-positive Cre-affected cell population in the mouse retinas were quantified by FACS and shown in the graph. Error bars, SD. Numbers of samples analyzed are shown in the graph (four independent litters). (C) RPCs that had progressed from S-phase to G2/M-phase for 3 hr (EdU;pH3-positive); PRs that had been born for 12 hr (EdU;Otx2-positive); and RPCs that had reentered cell cycle for 15 hr (CldU;IdU-positive) in the mouse retinas were counted and shown in the graph. Error bars denote SD (n = 5, five independent litters). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. (D) Relative mRNA levels of Hif1a, hexokinase 2 (Hk2), and pyruvate kinase M2 (Pkm2) in P0 mouse retinas with the indicated genotypes were examined by real-time quantitative polymerase chain reaction (RT-qPCR). The y-axis values are relative 2-ΔCt values against those of Tsc1fl/+;Tyrp1- Cre mouse retinas. Error bars, SD (n = 4, three independent litters). (E) Expressions of the indicated proteins in the mouse retinas were examined by Western blot (WB). Relative amounts of the proteins used in each sample were determined by WB detection of β-actin. (F) Relative WB band intensities of the proteins are determined by the ImageJ software and shown in the graph. Error bars, SD (n = 3, three independent litters). (G) Distributions of Pkm2, Tom20, and hypoxyprobe-labeled proteins in the mouse retinas were examined by immunostaining. (H) P0 Tsc1fl/fl;Tyrp1-Cre littermate mice were injected with a chemical inhibitor of glycolysis (2DG) or mitochondrial oxidative phosphorylation (Metformin), and the eye sections of the mice were

Journal: eLife

Article Title: mTORC1-induced retinal progenitor cell overproliferation leads to accelerated mitotic aging and degeneration of descendent Müller glia

doi: 10.7554/elife.70079

Figure Lengend Snippet: Figure 7. Hif1a supports mTORC1-induced retinal progenitor cell (RPC) proliferation through the expression of glycolytic enzymes. (A) The Cre-affected cells in P0 mouse retinas with the indicated genotypes were visualized by R26tdTom Cre reporter. Distribution of mTORC1-active cells in the boxed areas was also examined by immunostaining of pS6 and shown in the second row. Proliferation and cell cycle progression of RPCs in the mouse retinas were also examined by 5-ethynyl-2′-deoxyuridine (EdU)/5-chloro-2'-deoxyuridine (CldU) labeling and chasing experiments as described in Materials and methods. (B) R26tdTom-positive Cre-affected cell population in the mouse retinas were quantified by FACS and shown in the graph. Error bars, SD. Numbers of samples analyzed are shown in the graph (four independent litters). (C) RPCs that had progressed from S-phase to G2/M-phase for 3 hr (EdU;pH3-positive); PRs that had been born for 12 hr (EdU;Otx2-positive); and RPCs that had reentered cell cycle for 15 hr (CldU;IdU-positive) in the mouse retinas were counted and shown in the graph. Error bars denote SD (n = 5, five independent litters). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. (D) Relative mRNA levels of Hif1a, hexokinase 2 (Hk2), and pyruvate kinase M2 (Pkm2) in P0 mouse retinas with the indicated genotypes were examined by real-time quantitative polymerase chain reaction (RT-qPCR). The y-axis values are relative 2-ΔCt values against those of Tsc1fl/+;Tyrp1- Cre mouse retinas. Error bars, SD (n = 4, three independent litters). (E) Expressions of the indicated proteins in the mouse retinas were examined by Western blot (WB). Relative amounts of the proteins used in each sample were determined by WB detection of β-actin. (F) Relative WB band intensities of the proteins are determined by the ImageJ software and shown in the graph. Error bars, SD (n = 3, three independent litters). (G) Distributions of Pkm2, Tom20, and hypoxyprobe-labeled proteins in the mouse retinas were examined by immunostaining. (H) P0 Tsc1fl/fl;Tyrp1-Cre littermate mice were injected with a chemical inhibitor of glycolysis (2DG) or mitochondrial oxidative phosphorylation (Metformin), and the eye sections of the mice were

Article Snippet: 70079 18 of 27 Reagent type (species) or resource Designation Source or reference Identifiers Additional information Antibody Anti- BrdU(CldU) (Rat monoclonal) Novus Biologicals NB500- 169 IHC (1:200) Antibody Anti- BrdU(IdU) (Mouse monoclonal) EXBIO 11–286 C100 IHC (1:200) Antibody Anti- Brn3b (Rabbit polyclonal) Santa Cruz Biotechnology sc- 31989 IHC (1:200) Antibody Anti- Calbindin (Rabbit polyclonal) Swant Inc. CB- 38 IHC (1:200) Antibody Anti- Cdo (Goat polyclonal) R&D Systems AF2429 IHC (1:200) Antibody Anti- c- myc (Mouse monoclonal) Santa Cruz Biotechnology sc40 WB (1:1000) Antibody Anti- Cleaved caspase- 3 (Rabbit polyclonal) Cell Signaling Technology 9661 IHC (1:200) Antibody Anti- Ezrin (Mouse monoclonal) Invitrogen Biotechnology 35–7300 IHC (1:200) Antibody Anti- GFAP (Rabbit polyclonal) Abcam ab48050 IHC (1:200) Antibody Anti- GFP (Chicken polyclonal) Abcam ab13970 IHC (1:2000) Antibody Anti- Glutamine synthase (Rabbit polyclonal) Sigma- Aldrich G- 2781 IHC (1:200) Antibody Anti- Hif1a (Mouse monoclonal) R&D Systems MAB1536 WB (1:1000) Antibody Anti- Hif2α/Epas1 (Rabbit polyclonal) Novus Biologicals NB100- 122 WB (1:1000) Antibody Anti- Hk2 (Rabbit polyclonal) Cell Signaling Technology 2867 WB 1:1,000 IHC (1:200) Antibody Anti- Hydroxy- Hif1alpha (Rabbit polyclonal) Cell Signaling Technology 3434 WB (1:1000) Antibody Anti- M- opsin Merck Millipore AB5405 IHC (1:200) Antibody Anti- Otx2 (Rabbit polyclonal) Abcam ab25985 IHC (1:200) Antibody Anti- Otx2 (Rabbit polyclonal) Abcam ab183951 IHC (1:200) Antibody Anti- Otx2 (Goat polyclonal) R&D Systems AF1979- SP IHC (1:200) Antibody Anti- p21/Cip1 (Mouse monoclonal) Santa Cruz Biotechnology SC817 WB (1:1000) IHC (1:200) Antibody Anti- p53 (Mouse monoclonal) Merck Millipore OP03- 100 WB (1:1000) IHC (1:200) Antibody Anti- Pax6 (Rabbit polyclonal) Covance PRB- 278P IHC (1:200) Antibody Anti- Phospho Smad1/5(S463/465) (Rabbit polyclonal) Invitrogen Biotechnology 700047 IHC (1:200) Antibody Anti- phospho Smad2(ser465/467) (Rabbit polyclonal) - Cell Signaling Technology 18338 IHC (1:200) Antibody Anti- phospho- Histone H3(S10) (pH3; Rabbit polyclonal) Merck Millipore 04–1093 IHC (1:200) Antibody Anti- phospho- S6(S235/236) (pS6; Rabbit polyclonal) Cell Signaling Technology 2211 WB (1:1000) IHC (1:200) Continued Continued on next page Lim et al. eLife 2021;10:e70079.

Techniques: Expressing, Immunostaining, Labeling, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Software, Injection, Phospho-proteomics

Fig. 4. Signal pathway identification and transcription factor activity verification. A: Representative western blotting strips of PAs from MCT-induced PAH rats with or without metformin treatment and densitometric analysis showing Hif1α, PKG relative to β-actin; B: Representative western blotting strips of PAs from MCT- induced PAH rats with or without metformin treatment and densitometric analysis showing p-AKT relative to AKT; C: Representative western blotting strips of PAs from MCT-induced PAH rats with or without metformin treatment and densitometric analysis showing p53 relative to β-actin; D: Representative western blotting strips of PAs from MCT-induced PAH rats with or without metformin treatment and densitometric analysis showing Sp1, Est1 relative to β-actin. MCT means monocrotaline; Met means metformin. **p < 0.01. n = 3.

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Long non-coding RNA and mRNA profile analysis of metformin to reverse the pulmonary hypertension vascular remodeling induced by monocrotaline.

doi: 10.1016/j.biopha.2019.108933

Figure Lengend Snippet: Fig. 4. Signal pathway identification and transcription factor activity verification. A: Representative western blotting strips of PAs from MCT-induced PAH rats with or without metformin treatment and densitometric analysis showing Hif1α, PKG relative to β-actin; B: Representative western blotting strips of PAs from MCT- induced PAH rats with or without metformin treatment and densitometric analysis showing p-AKT relative to AKT; C: Representative western blotting strips of PAs from MCT-induced PAH rats with or without metformin treatment and densitometric analysis showing p53 relative to β-actin; D: Representative western blotting strips of PAs from MCT-induced PAH rats with or without metformin treatment and densitometric analysis showing Sp1, Est1 relative to β-actin. MCT means monocrotaline; Met means metformin. **p < 0.01. n = 3.

Article Snippet: The dilution ratio of all primary antibodies used was as follows: (PKG, catalog number: 21646-1-AP, 1:1000 dilution, proteintech), (Sp1, catalog number: 21962-1-AP, 1:2000 dilution, proteintech), (Est1, catalog number: 12522-1-AP, 1:1000 dilution, proteintech), (Hif1α, Cat. #: AF1009, 1:2000 dilution, Affinity Biosciences), (p53, catalog number: 10442-1-AP, 1:5000 dilution, proteintech), (Cyclin D1, catalog number: 26755-1-AP, 1:1000 dilution, proteintech), (β-actin, catalog number: 60008-1-1 g, 1:10000 dilution, proteintech), (AKT, catalog number: 10176-2-AP, 1:2000 dilution, proteintech), (p-AKT, # 9271, 1:1000 dilution, Cell Signaling Technology).

Techniques: Activity Assay, Western Blot

Fig. 7. Mechanism analysis of NONRATT015587.2 regulation of pulmonary vascular remodeling in vitro assay and differential gene clustergram in MCT-induced PAH rats and controls, or metformin treatment. A-G: Realtime-PCR analysis of p53, Est1, Rela, Cebpa/b, Sp1, Hif1α expression in pcDNA, pcDNA- NONRATT015587.2, and pcDNA- NONRATT015587.2+metformin group. H: Representative western blotting strips of PASMCs from pcDNA-NONRATT015587.2 with or without metformin treatment and densitometric analysis showing Hif1α, p53 relative to β-actin; I: The differential expression of mRNA in the microarray analysis. Hierarchical clustering analysis is shown for 32 mRNA (fold change > 2, p < 0.05) in the control, monocrotaline and monocrotaline + metformin group using gene microarray analysis. MCT means monocrotaline; Met means metformin. **p < 0.01. n = 3.

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Long non-coding RNA and mRNA profile analysis of metformin to reverse the pulmonary hypertension vascular remodeling induced by monocrotaline.

doi: 10.1016/j.biopha.2019.108933

Figure Lengend Snippet: Fig. 7. Mechanism analysis of NONRATT015587.2 regulation of pulmonary vascular remodeling in vitro assay and differential gene clustergram in MCT-induced PAH rats and controls, or metformin treatment. A-G: Realtime-PCR analysis of p53, Est1, Rela, Cebpa/b, Sp1, Hif1α expression in pcDNA, pcDNA- NONRATT015587.2, and pcDNA- NONRATT015587.2+metformin group. H: Representative western blotting strips of PASMCs from pcDNA-NONRATT015587.2 with or without metformin treatment and densitometric analysis showing Hif1α, p53 relative to β-actin; I: The differential expression of mRNA in the microarray analysis. Hierarchical clustering analysis is shown for 32 mRNA (fold change > 2, p < 0.05) in the control, monocrotaline and monocrotaline + metformin group using gene microarray analysis. MCT means monocrotaline; Met means metformin. **p < 0.01. n = 3.

Article Snippet: The dilution ratio of all primary antibodies used was as follows: (PKG, catalog number: 21646-1-AP, 1:1000 dilution, proteintech), (Sp1, catalog number: 21962-1-AP, 1:2000 dilution, proteintech), (Est1, catalog number: 12522-1-AP, 1:1000 dilution, proteintech), (Hif1α, Cat. #: AF1009, 1:2000 dilution, Affinity Biosciences), (p53, catalog number: 10442-1-AP, 1:5000 dilution, proteintech), (Cyclin D1, catalog number: 26755-1-AP, 1:1000 dilution, proteintech), (β-actin, catalog number: 60008-1-1 g, 1:10000 dilution, proteintech), (AKT, catalog number: 10176-2-AP, 1:2000 dilution, proteintech), (p-AKT, # 9271, 1:1000 dilution, Cell Signaling Technology).

Techniques: In Vitro, Expressing, Western Blot, Quantitative Proteomics, Microarray, Control