Journal: Scientific Reports

Article Title: Heat shock protein 22 alleviates doxorubicin-induced kidney injury by suppressing oxidative stress and apoptosis

doi: 10.1038/s41598-024-75277-5

Figure Lengend Snippet: A An overview of the animal treatment protocols conducted in the present study. B and C the protein expression of HSP22 was investigated by western blotting in the kidney tissues ( n = 6). D IHC staining was conducted to evaluate the protein expression of HSP22 in the kidney tissues ( n = 6). E and F The serum levels of SCr and BUN were detected ( n = 6). G H&E staining was conducted for evaluating the pathological changes in the kidney tissues ( n = 6). ns means no statistically significant, ** p < 0.01 and *** p < 0.001. Data represent the mean ± SD of at least 3 times of separate experiments.

Article Snippet: The following primary antibodies were used: anti-HSP22 (CAT NO. ab151552; 1:1000; Abcam, Cambridge, MA, USA), anti-NOX-2 (CAT NO. ab310337; 1:1000; Abcam), anti-NOX-4 (CAT NO. ab154244; 1:1000; Abcam), anti-SOD-1 (CAT NO. CST-65778SF; 1:1000; Cell Signaling Technology, Danvers, MA, USA), anti-SOD-2 (CAT NO. CST-13141 S; 1:1000; Cell Signaling Technology), anti-Bax (CAT NO. ab32503; 1:1000; Abcam), anti-Bcl-2 (CAT NO. ab182858; 1:1000; Abcam), anti-Nrf2 (CAT NO. ab92946; 1:1000; Abcam), and anti-HO-1 (CAT NO. ab52947; 1:1000; Abcam).

Techniques: Expressing, Western Blot, Immunohistochemistry, Staining

Journal: Scientific Reports

Article Title: Heat shock protein 22 alleviates doxorubicin-induced kidney injury by suppressing oxidative stress and apoptosis

doi: 10.1038/s41598-024-75277-5

Figure Lengend Snippet: A and B the protein expression of HSP22 was investigated by western blotting in HK2 cells ( n = 6). C – E , the expressions of pro-oxidative stress proteins, such as NOX-2 and NOX-4, were detected by western blotting in HK2 cells ( n = 6). C , F , and G the expressions of antioxidant proteins, such as SOD-1 and SOD-2, were detected by western blotting in HK2 cells ( n = 6). H The flow cytometry assay was conducted to evaluate apoptotic rates in different groups of HK2 cells ( n = 6). I , J , and K The protein expressions of pro-apoptosis protein Bax and anti-apoptosis protein Bcl-2 were investigated by western blotting in HK2 cells ( n = 6). L , M , and N The protein expressions of Nrf2 and HO-1 were investigated by western blotting in HK2 cells ( n = 6). ns means no statistically significant, * p < 0.05 and ** p < 0.01 and *** p < 0.001. Data represent the mean ± SD of at least 3 times of separate experiments.

Article Snippet: The following primary antibodies were used: anti-HSP22 (CAT NO. ab151552; 1:1000; Abcam, Cambridge, MA, USA), anti-NOX-2 (CAT NO. ab310337; 1:1000; Abcam), anti-NOX-4 (CAT NO. ab154244; 1:1000; Abcam), anti-SOD-1 (CAT NO. CST-65778SF; 1:1000; Cell Signaling Technology, Danvers, MA, USA), anti-SOD-2 (CAT NO. CST-13141 S; 1:1000; Cell Signaling Technology), anti-Bax (CAT NO. ab32503; 1:1000; Abcam), anti-Bcl-2 (CAT NO. ab182858; 1:1000; Abcam), anti-Nrf2 (CAT NO. ab92946; 1:1000; Abcam), and anti-HO-1 (CAT NO. ab52947; 1:1000; Abcam).

Techniques: Expressing, Western Blot, Flow Cytometry

Journal: Scientific Reports

Article Title: Heat shock protein 22 alleviates doxorubicin-induced kidney injury by suppressing oxidative stress and apoptosis

doi: 10.1038/s41598-024-75277-5

Figure Lengend Snippet: A and B the protein expression of Nrf2 was investigated by western blotting in HK2 cells ( n = 6). C – E , the expressions of HSP22 and HO-1 were detected by western blotting in HK2 cells ( n = 6). C , F , and G the expressions of pro-oxidative stress proteins, such as NOX-2 and NOX-4, were detected by western blotting in HK2 cells ( n = 6). C , H , and I the expressions of antioxidant proteins, such as SOD-1 and SOD-2, were detected by western blotting in HK2 cells ( n = 6). J , K , and L the protein expressions of pro-apoptosis protein Bax and anti-apoptosis protein Bcl-2 were investigated by western blotting in HK2 cells ( n = 6). ns means no statistically significant, * p < 0.05, ** p < 0.01 and *** p < 0.001. Data represent the mean ± SD of at least 3 times of separate experiments.

Article Snippet: The following primary antibodies were used: anti-HSP22 (CAT NO. ab151552; 1:1000; Abcam, Cambridge, MA, USA), anti-NOX-2 (CAT NO. ab310337; 1:1000; Abcam), anti-NOX-4 (CAT NO. ab154244; 1:1000; Abcam), anti-SOD-1 (CAT NO. CST-65778SF; 1:1000; Cell Signaling Technology, Danvers, MA, USA), anti-SOD-2 (CAT NO. CST-13141 S; 1:1000; Cell Signaling Technology), anti-Bax (CAT NO. ab32503; 1:1000; Abcam), anti-Bcl-2 (CAT NO. ab182858; 1:1000; Abcam), anti-Nrf2 (CAT NO. ab92946; 1:1000; Abcam), and anti-HO-1 (CAT NO. ab52947; 1:1000; Abcam).

Techniques: Expressing, Western Blot

Journal: Scientific Reports

Article Title: Heat shock protein 22 alleviates doxorubicin-induced kidney injury by suppressing oxidative stress and apoptosis

doi: 10.1038/s41598-024-75277-5

Figure Lengend Snippet: Graphical abstract of the effects of HSP22 on DOX-induced kidney injury. HSP22 alleviates DOX-induced kidney injury by inhibiting oxidative stress and apoptosis, and the molecular mechanism involved activation of the Nrf2/HO-1-signaling pathway.

Article Snippet: The following primary antibodies were used: anti-HSP22 (CAT NO. ab151552; 1:1000; Abcam, Cambridge, MA, USA), anti-NOX-2 (CAT NO. ab310337; 1:1000; Abcam), anti-NOX-4 (CAT NO. ab154244; 1:1000; Abcam), anti-SOD-1 (CAT NO. CST-65778SF; 1:1000; Cell Signaling Technology, Danvers, MA, USA), anti-SOD-2 (CAT NO. CST-13141 S; 1:1000; Cell Signaling Technology), anti-Bax (CAT NO. ab32503; 1:1000; Abcam), anti-Bcl-2 (CAT NO. ab182858; 1:1000; Abcam), anti-Nrf2 (CAT NO. ab92946; 1:1000; Abcam), and anti-HO-1 (CAT NO. ab52947; 1:1000; Abcam).

Techniques: Activation Assay

Journal: Frontiers in Veterinary Science

Article Title: Feline coronavirus influences the biogenesis and composition of extracellular vesicles derived from CRFK cells

doi: 10.3389/fvets.2024.1388438

Figure Lengend Snippet: The influence of FCoV infection on transmembrane molecules, caspases, and initiation of stress-specific responses in response to FCoV infection.

Article Snippet: Then, the membrane was washed three times for 10 min each with 1× Tris-buffered saline containing Tween-20 (0.2%) buffer solution (TBST) and incubated with primary antibodies including Alix (Fisher Scientific), CD63 (Santa Cruz Biotechnology), TSG101 (Fisher Scientific), ACE2 (DHSB), TMPRSS2 (DHSB), anti-flotillin-1 (BD Bioscience), Clathrin (BD Bioscience), cadherin (DSHB), CD29 (DSHB), anti-TLR3 (Abnova), TLR6 and 7 (Invitrogen), IRF4 (DSHB), mCCL22 (RD Systems), TGFβ-3(DHSB), TNF-α (Bioss Antibodies Inc.), CD47(Bioss Antibodies Inc.), LAMP-1 (human) (DSHB), ATPase (DSHB), TSPAN8, HSPB8-13B6 (Hsp22) (Invitrogen), HSPB1-1 (Hsp27) (DSHB), Hsp100 (DSHB), DIS3-1D7 (DSHB), and cleaved caspase-3 (RD Systems).

Techniques: Infection, Activation Assay

Journal: Frontiers in Veterinary Science

Article Title: Feline coronavirus influences the biogenesis and composition of extracellular vesicles derived from CRFK cells

doi: 10.3389/fvets.2024.1388438

Figure Lengend Snippet: The influence of FCoV infection on transmembrane molecules, caspases, and initiation of stress-specific responses in response to FCoV infection.

Article Snippet: Then, the membrane was washed three times for 10 min each with 1× Tris-buffered saline containing Tween-20 (0.2%) buffer solution (TBST) and incubated with primary antibodies including Alix (Fisher Scientific), CD63 (Santa Cruz Biotechnology), TSG101 (Fisher Scientific), ACE2 (DHSB), TMPRSS2 (DHSB), anti-flotillin-1 (BD Bioscience), Clathrin (BD Bioscience), cadherin (DSHB), CD29 (DSHB), anti-TLR3 (Abnova), TLR6 and 7 (Invitrogen), IRF4 (DSHB), mCCL22 (RD Systems), TGFβ-3(DHSB), TNF-α (Bioss Antibodies Inc.), CD47(Bioss Antibodies Inc.), LAMP-1 (human) (DSHB), ATPase (DSHB), TSPAN8, HSPB8-13B6 (Hsp22) (Invitrogen), HSPB1-1 (Hsp27) (DSHB), Hsp100 (DSHB), DIS3-1D7 (DSHB), and cleaved caspase-3 (RD Systems).

Techniques: Infection, Activation Assay

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet: Primers used for qRT–PCR analysis

Article Snippet: Rabbit monoclonal anti HSP22 , Abcam , ab151552.

Techniques:

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet: RSG improves diabetes-induced vascular endothelial injury and increases HSP22 expression in vivo (A) Blood glucose levels of mice in the control, T2DM and T2DM + RSG groups were measured at baseline and at 4, 8, and 12 weeks. (B) The body weights (BWs) of mice in the control, T2DM and T2DM + RSG groups were measured at baseline and at 4, 8, and 12 weeks. (C) Representative images of hematoxylin and eosin staining (H&E, 40× magnification). (D) Representative immunohistochemical staining images of ICAM-1, PPAR-γ and HSP22 in vascular sections (40× magnification). (E) Quantification of positive staining for ICAM-1 in vascular rings. (F) Quantification of positive staining for PPAR-γ in vascular rings. (G) Quantification of positive staining for HSP22 in vascular sections. (H) mRNA expression of IL5, IL6, IL13, Mip3α and TGFβ-1 in vascular tissue. Mean ± SEM two-way ANOVA followed by Tukey’s post hoc. ∗ p< 0.05 T2DM group compared to control group; # p< 0.05 T2DM group compared to T2DM + RSG group. Scale bar 20μm.

Article Snippet: Rabbit monoclonal anti HSP22 , Abcam , ab151552.

Techniques: Expressing, In Vivo, Staining, Immunohistochemical staining

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet: RSG inhibits hyperglycemia-induced endothelial activation and upregulates HSP22 expression in vitro (A) Cell viability was evaluated in cells in 96-well plates by a Cell Counting Kit-8 (CCK8) after treatment with NG, HG and HG + RSG for 24 h. (B) Cytotoxicity was measured in cells in 96-well plates by lactate dehydrogenase (LDH) release after treatment with NG, HG and HG + RSG for 24 h. (C) Representative images of the adhesion of primary human peripheral mononuclear cells (PBMCs) to stimulated HUVECs. (D) Quantification of the monocyte adhesion assay. (E) mRNA expression of ICAM-1 was determined by qRT–PCR analysis. (F) Endothelial cell activation-related cytokines were examined by qRT–PCR analysis. (G) PPAR-γ and HSP22 protein expression levels were measured by Western blot analysis. (H) Quantification of PPAR-γ protein expression. (I) Quantification of HSP22 protein expression. GAPDH was used as a loading control. Mean ± SEM two-way ANOVA followed by Tukey’s post hoc. ∗ p< 0.05 HG group compared to NG group; # p< 0.05 HG group compared to HG + RSG group. Scale bar 100μm.

Article Snippet: Rabbit monoclonal anti HSP22 , Abcam , ab151552.

Techniques: Activation Assay, Expressing, In Vitro, Cell Counting, Cell Adhesion Assay, Quantitative RT-PCR, Western Blot

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet: RSG improves diabetes-induced vascular endothelial injury partially via HSP22 expression in vivo (A) Blood glucose levels of mice in the WT + T2DM + RSG and HSP22-KO + T2DM + RSG groups were measured at baseline and 4, 8, and 12 weeks. (B) The body weights (BWs) of mice in the WT + T2DM + RSG and HSP22-KO + T2DM + RSG groups were measured at baseline and at 4, 8, and 12 weeks. (C) Representative images of H&E staining (40× magnification). (D) Representative immunohistochemical staining images of PPAR-γ, HSP22 and ICAM-1 in vascular sections (40× magnification). (E) Quantification of positive staining for PPAR-γ in vascular rings. (F) Quantification of positive staining for HSP22 in vascular rings. (G) Quantification of positive staining for ICAM-1 in vascular sections. (H) mRNA expression of IL5, IL6, IL13, Mip3α and TGFβ-1 in vascular tissue. Mean ± SEM t tests. ∗ p< 0.05 HSP22-KO + T2DM + RSG group compared to WT + T2DM + RSG group. Scale bar 20μm.

Article Snippet: Rabbit monoclonal anti HSP22 , Abcam , ab151552.

Techniques: Expressing, In Vivo, Staining, Immunohistochemical staining

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet: RSG inhibits hyperglycemia-induced endothelial activation partially via HSP22 expression in vitro (A) PPAR-γ and HSP22 protein expression levels were determined by Western blot analysis. (B) Quantification of PPAR-γ protein expression. (C) Quantification of HSP22 protein expression. GAPDH was used as a loading control. (D) Cell viability was evaluated in cells in 96-well plates by CCK8 assay. (E) Cytotoxicity was measured in cells in 96-well plates by LDH release. (F) Representative images of the adhesion of PBMCs to stimulated HUVECs. (G) Quantification of the monocyte adhesion assay. (H) mRNA expression of ICAM-1 was determined by RT–PCR analysis. (I) Endothelial cell activation-related cytokines were examined by RT–PCR analysis. Mean ± SEM t tests. ∗ p< 0.05 HSP22 si + HG + RSG group compared to NC si + HG + RSG group. Scale bar 100μm.

Article Snippet: Rabbit monoclonal anti HSP22 , Abcam , ab151552.

Techniques: Activation Assay, Expressing, In Vitro, Western Blot, CCK-8 Assay, Cell Adhesion Assay, Reverse Transcription Polymerase Chain Reaction

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet: RSG alleviates diabetes-induced oxidative stress and mitochondrial impairment partially via HSP22 expression in vivo (A) Representative images of DHE staining in vascular sections (40× magnification). (B) Quantification of the fluorescence intensity of ROS levels in vascular sections. (C) Quantification of 8-OHdG levels in vascular sections. Mean ± SEM t tests. ∗ p< 0.05 HSP22-KO + T2DM + RSG group compared to WT + T2DM + RSG group. Scale bar 20μm.

Article Snippet: Rabbit monoclonal anti HSP22 , Abcam , ab151552.

Techniques: Expressing, In Vivo, Staining, Fluorescence

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet: RSG inhibits hyperglycemia-induced mitochondrial oxidative stress and dysfunction in a manner partially dependent on HSP22 expression in vitro (A) Quantification of mitochondrial superoxide levels in HUVECs by flow cytometry. (B) Representative images of MitoTracker Red staining showing the mitochondrial morphology and density in HUVECs (63× magnification). (C) Quantification of mitochondrial density in HUVECs. (D) Representative images of JC-1 immunochemistry staining showing the mitochondrial membrane potential in HUVECs (63× magnification). (E) Quantification of mitochondrial membrane potential in HUVECs. (F) mRNA expression of SOD2 was determined by RT–PCR analysis. (G) ATP levels were measured in cells in 96-well plates by an ATP Kit after treatment with NG, HG and HG + RSG for 24 h. Mean ± SEM t tests. ∗ p< 0.05 HSP22 si + HG + RSG group compared to NC si + HG + RSG group. Scale bar 20μm.

Article Snippet: Rabbit monoclonal anti HSP22 , Abcam , ab151552.

Techniques: Expressing, In Vitro, Flow Cytometry, Staining, Reverse Transcription Polymerase Chain Reaction

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti HSP22 , Abcam , ab151552.

Techniques: Recombinant, CCK-8 Assay, Lysis, Bicinchoninic Acid Protein Assay, Sequencing, Software, Confocal Laser Scanning Microscopy

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet: Primers used for qRT–PCR analysis

Article Snippet: The membranes were incubated with the following primary antibodies: HSP22 antibody (Abcam, ab151552, rabbit monoclonal, 1:200, USA), PPAR-γ antibody (CST, 2435, rabbit polyclonal, 1:500, USA) and GAPDH antibody (Abcam, ab8245, mouse monoclonal, 1:2000, USA).

Techniques:

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet: RSG improves diabetes-induced vascular endothelial injury and increases HSP22 expression in vivo (A) Blood glucose levels of mice in the control, T2DM and T2DM + RSG groups were measured at baseline and at 4, 8, and 12 weeks. (B) The body weights (BWs) of mice in the control, T2DM and T2DM + RSG groups were measured at baseline and at 4, 8, and 12 weeks. (C) Representative images of hematoxylin and eosin staining (H&E, 40× magnification). (D) Representative immunohistochemical staining images of ICAM-1, PPAR-γ and HSP22 in vascular sections (40× magnification). (E) Quantification of positive staining for ICAM-1 in vascular rings. (F) Quantification of positive staining for PPAR-γ in vascular rings. (G) Quantification of positive staining for HSP22 in vascular sections. (H) mRNA expression of IL5, IL6, IL13, Mip3α and TGFβ-1 in vascular tissue. Mean ± SEM two-way ANOVA followed by Tukey’s post hoc. ∗ p< 0.05 T2DM group compared to control group; # p< 0.05 T2DM group compared to T2DM + RSG group. Scale bar 20μm.

Article Snippet: The membranes were incubated with the following primary antibodies: HSP22 antibody (Abcam, ab151552, rabbit monoclonal, 1:200, USA), PPAR-γ antibody (CST, 2435, rabbit polyclonal, 1:500, USA) and GAPDH antibody (Abcam, ab8245, mouse monoclonal, 1:2000, USA).

Techniques: Expressing, In Vivo, Staining, Immunohistochemical staining

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet: RSG inhibits hyperglycemia-induced endothelial activation and upregulates HSP22 expression in vitro (A) Cell viability was evaluated in cells in 96-well plates by a Cell Counting Kit-8 (CCK8) after treatment with NG, HG and HG + RSG for 24 h. (B) Cytotoxicity was measured in cells in 96-well plates by lactate dehydrogenase (LDH) release after treatment with NG, HG and HG + RSG for 24 h. (C) Representative images of the adhesion of primary human peripheral mononuclear cells (PBMCs) to stimulated HUVECs. (D) Quantification of the monocyte adhesion assay. (E) mRNA expression of ICAM-1 was determined by qRT–PCR analysis. (F) Endothelial cell activation-related cytokines were examined by qRT–PCR analysis. (G) PPAR-γ and HSP22 protein expression levels were measured by Western blot analysis. (H) Quantification of PPAR-γ protein expression. (I) Quantification of HSP22 protein expression. GAPDH was used as a loading control. Mean ± SEM two-way ANOVA followed by Tukey’s post hoc. ∗ p< 0.05 HG group compared to NG group; # p< 0.05 HG group compared to HG + RSG group. Scale bar 100μm.

Article Snippet: The membranes were incubated with the following primary antibodies: HSP22 antibody (Abcam, ab151552, rabbit monoclonal, 1:200, USA), PPAR-γ antibody (CST, 2435, rabbit polyclonal, 1:500, USA) and GAPDH antibody (Abcam, ab8245, mouse monoclonal, 1:2000, USA).

Techniques: Activation Assay, Expressing, In Vitro, Cell Counting, Cell Adhesion Assay, Quantitative RT-PCR, Western Blot

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet: RSG improves diabetes-induced vascular endothelial injury partially via HSP22 expression in vivo (A) Blood glucose levels of mice in the WT + T2DM + RSG and HSP22-KO + T2DM + RSG groups were measured at baseline and 4, 8, and 12 weeks. (B) The body weights (BWs) of mice in the WT + T2DM + RSG and HSP22-KO + T2DM + RSG groups were measured at baseline and at 4, 8, and 12 weeks. (C) Representative images of H&E staining (40× magnification). (D) Representative immunohistochemical staining images of PPAR-γ, HSP22 and ICAM-1 in vascular sections (40× magnification). (E) Quantification of positive staining for PPAR-γ in vascular rings. (F) Quantification of positive staining for HSP22 in vascular rings. (G) Quantification of positive staining for ICAM-1 in vascular sections. (H) mRNA expression of IL5, IL6, IL13, Mip3α and TGFβ-1 in vascular tissue. Mean ± SEM t tests. ∗ p< 0.05 HSP22-KO + T2DM + RSG group compared to WT + T2DM + RSG group. Scale bar 20μm.

Article Snippet: The membranes were incubated with the following primary antibodies: HSP22 antibody (Abcam, ab151552, rabbit monoclonal, 1:200, USA), PPAR-γ antibody (CST, 2435, rabbit polyclonal, 1:500, USA) and GAPDH antibody (Abcam, ab8245, mouse monoclonal, 1:2000, USA).

Techniques: Expressing, In Vivo, Staining, Immunohistochemical staining

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet: RSG inhibits hyperglycemia-induced endothelial activation partially via HSP22 expression in vitro (A) PPAR-γ and HSP22 protein expression levels were determined by Western blot analysis. (B) Quantification of PPAR-γ protein expression. (C) Quantification of HSP22 protein expression. GAPDH was used as a loading control. (D) Cell viability was evaluated in cells in 96-well plates by CCK8 assay. (E) Cytotoxicity was measured in cells in 96-well plates by LDH release. (F) Representative images of the adhesion of PBMCs to stimulated HUVECs. (G) Quantification of the monocyte adhesion assay. (H) mRNA expression of ICAM-1 was determined by RT–PCR analysis. (I) Endothelial cell activation-related cytokines were examined by RT–PCR analysis. Mean ± SEM t tests. ∗ p< 0.05 HSP22 si + HG + RSG group compared to NC si + HG + RSG group. Scale bar 100μm.

Article Snippet: The membranes were incubated with the following primary antibodies: HSP22 antibody (Abcam, ab151552, rabbit monoclonal, 1:200, USA), PPAR-γ antibody (CST, 2435, rabbit polyclonal, 1:500, USA) and GAPDH antibody (Abcam, ab8245, mouse monoclonal, 1:2000, USA).

Techniques: Activation Assay, Expressing, In Vitro, Western Blot, CCK-8 Assay, Cell Adhesion Assay, Reverse Transcription Polymerase Chain Reaction

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet: RSG alleviates diabetes-induced oxidative stress and mitochondrial impairment partially via HSP22 expression in vivo (A) Representative images of DHE staining in vascular sections (40× magnification). (B) Quantification of the fluorescence intensity of ROS levels in vascular sections. (C) Quantification of 8-OHdG levels in vascular sections. Mean ± SEM t tests. ∗ p< 0.05 HSP22-KO + T2DM + RSG group compared to WT + T2DM + RSG group. Scale bar 20μm.

Article Snippet: The membranes were incubated with the following primary antibodies: HSP22 antibody (Abcam, ab151552, rabbit monoclonal, 1:200, USA), PPAR-γ antibody (CST, 2435, rabbit polyclonal, 1:500, USA) and GAPDH antibody (Abcam, ab8245, mouse monoclonal, 1:2000, USA).

Techniques: Expressing, In Vivo, Staining, Fluorescence

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet: RSG inhibits hyperglycemia-induced mitochondrial oxidative stress and dysfunction in a manner partially dependent on HSP22 expression in vitro (A) Quantification of mitochondrial superoxide levels in HUVECs by flow cytometry. (B) Representative images of MitoTracker Red staining showing the mitochondrial morphology and density in HUVECs (63× magnification). (C) Quantification of mitochondrial density in HUVECs. (D) Representative images of JC-1 immunochemistry staining showing the mitochondrial membrane potential in HUVECs (63× magnification). (E) Quantification of mitochondrial membrane potential in HUVECs. (F) mRNA expression of SOD2 was determined by RT–PCR analysis. (G) ATP levels were measured in cells in 96-well plates by an ATP Kit after treatment with NG, HG and HG + RSG for 24 h. Mean ± SEM t tests. ∗ p< 0.05 HSP22 si + HG + RSG group compared to NC si + HG + RSG group. Scale bar 20μm.

Article Snippet: The membranes were incubated with the following primary antibodies: HSP22 antibody (Abcam, ab151552, rabbit monoclonal, 1:200, USA), PPAR-γ antibody (CST, 2435, rabbit polyclonal, 1:500, USA) and GAPDH antibody (Abcam, ab8245, mouse monoclonal, 1:2000, USA).

Techniques: Expressing, In Vitro, Flow Cytometry, Staining, Reverse Transcription Polymerase Chain Reaction

Journal: iScience

Article Title: Rosiglitazone reduces diabetes angiopathy by inhibiting mitochondrial dysfunction dependent on regulating HSP22 expression

doi: 10.1016/j.isci.2023.106194

Figure Lengend Snippet:

Article Snippet: The membranes were incubated with the following primary antibodies: HSP22 antibody (Abcam, ab151552, rabbit monoclonal, 1:200, USA), PPAR-γ antibody (CST, 2435, rabbit polyclonal, 1:500, USA) and GAPDH antibody (Abcam, ab8245, mouse monoclonal, 1:2000, USA).

Techniques: Recombinant, CCK-8 Assay, Lysis, Bicinchoninic Acid Protein Assay, Sequencing, Software, Confocal Laser Scanning Microscopy