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Neuroprotection by paricalcitol in vivo is VDR-dependent and mediated by the cAMP-PKA-DRP1 pathway. (A, B) Representative Western blot bands and quantitative analysis for VDR, ACSL4, and 4HNE after VDR siRNA administration in each group. (C) Results of the modified Garcia test, forelimb placement test, and corner turn test at 24 ​h post-ICH after VDR siRNA administration (n ​= ​6/group). (D) Representative Western blot bands for cAMP, p-PKA, PKA, p-DRP1 (Ser637), ACSL4, and 4HNE at 24 ​h post-ICH with <t>SQ22536</t> injection. (E) Quantitative analysis of cAMP, p-PKA, PKA, p-DRP1 (Ser637), ACSL4, and 4HNE levels. (F) Results of the modified Garcia test, forelimb placement test, and corner turn test after SQ22536 administration. (G, H) Quantitative ELISA analysis of brain MDA and GSH level after SQ22536 administration. Data are presented as mean ​± ​SD. ∗p ​< ​0.05 vs. sham group; @p ​< ​0.05 vs. ICH ​+ ​vehicle group; &p ​< ​0.05 vs. ICH ​+ ​PAL ​+ ​DMSO group. (n ​= ​6/group). #p ​< ​0.05 vs ICH ​+ ​Vehicle ​+ ​Scr siRNA group; $p ​< ​0.05 vs ICH ​+ ​PAL ​+ ​Scr siRNA group; ∗p ​< ​0.05 vs. sham group; @p ​< ​0.05 vs. ICH ​+ ​vehicle group; &p ​< ​0.05 vs. ICH ​+ ​PAL ​+ ​DMSO group. (n ​= ​6/group).
Camp Inhibitor Sq22536, supplied by Tocris, 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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adenylyl cyclase sq 22536  (Tocris)
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Neuroprotection by paricalcitol in vivo is VDR-dependent and mediated by the cAMP-PKA-DRP1 pathway. (A, B) Representative Western blot bands and quantitative analysis for VDR, ACSL4, and 4HNE after VDR siRNA administration in each group. (C) Results of the modified Garcia test, forelimb placement test, and corner turn test at 24 ​h post-ICH after VDR siRNA administration (n ​= ​6/group). (D) Representative Western blot bands for cAMP, p-PKA, PKA, p-DRP1 (Ser637), ACSL4, and 4HNE at 24 ​h post-ICH with <t>SQ22536</t> injection. (E) Quantitative analysis of cAMP, p-PKA, PKA, p-DRP1 (Ser637), ACSL4, and 4HNE levels. (F) Results of the modified Garcia test, forelimb placement test, and corner turn test after SQ22536 administration. (G, H) Quantitative ELISA analysis of brain MDA and GSH level after SQ22536 administration. Data are presented as mean ​± ​SD. ∗p ​< ​0.05 vs. sham group; @p ​< ​0.05 vs. ICH ​+ ​vehicle group; &p ​< ​0.05 vs. ICH ​+ ​PAL ​+ ​DMSO group. (n ​= ​6/group). #p ​< ​0.05 vs ICH ​+ ​Vehicle ​+ ​Scr siRNA group; $p ​< ​0.05 vs ICH ​+ ​PAL ​+ ​Scr siRNA group; ∗p ​< ​0.05 vs. sham group; @p ​< ​0.05 vs. ICH ​+ ​vehicle group; &p ​< ​0.05 vs. ICH ​+ ​PAL ​+ ​DMSO group. (n ​= ​6/group).
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Neuroprotection by paricalcitol in vivo is VDR-dependent and mediated by the cAMP-PKA-DRP1 pathway. (A, B) Representative Western blot bands and quantitative analysis for VDR, ACSL4, and 4HNE after VDR siRNA administration in each group. (C) Results of the modified Garcia test, forelimb placement test, and corner turn test at 24 ​h post-ICH after VDR siRNA administration (n ​= ​6/group). (D) Representative Western blot bands for cAMP, p-PKA, PKA, p-DRP1 (Ser637), ACSL4, and 4HNE at 24 ​h post-ICH with <t>SQ22536</t> injection. (E) Quantitative analysis of cAMP, p-PKA, PKA, p-DRP1 (Ser637), ACSL4, and 4HNE levels. (F) Results of the modified Garcia test, forelimb placement test, and corner turn test after SQ22536 administration. (G, H) Quantitative ELISA analysis of brain MDA and GSH level after SQ22536 administration. Data are presented as mean ​± ​SD. ∗p ​< ​0.05 vs. sham group; @p ​< ​0.05 vs. ICH ​+ ​vehicle group; &p ​< ​0.05 vs. ICH ​+ ​PAL ​+ ​DMSO group. (n ​= ​6/group). #p ​< ​0.05 vs ICH ​+ ​Vehicle ​+ ​Scr siRNA group; $p ​< ​0.05 vs ICH ​+ ​PAL ​+ ​Scr siRNA group; ∗p ​< ​0.05 vs. sham group; @p ​< ​0.05 vs. ICH ​+ ​vehicle group; &p ​< ​0.05 vs. ICH ​+ ​PAL ​+ ​DMSO group. (n ​= ​6/group).
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Neuroprotection by paricalcitol in vivo is VDR-dependent and mediated by the cAMP-PKA-DRP1 pathway. (A, B) Representative Western blot bands and quantitative analysis for VDR, ACSL4, and 4HNE after VDR siRNA administration in each group. (C) Results of the modified Garcia test, forelimb placement test, and corner turn test at 24 ​h post-ICH after VDR siRNA administration (n ​= ​6/group). (D) Representative Western blot bands for cAMP, p-PKA, PKA, p-DRP1 (Ser637), ACSL4, and 4HNE at 24 ​h post-ICH with <t>SQ22536</t> injection. (E) Quantitative analysis of cAMP, p-PKA, PKA, p-DRP1 (Ser637), ACSL4, and 4HNE levels. (F) Results of the modified Garcia test, forelimb placement test, and corner turn test after SQ22536 administration. (G, H) Quantitative ELISA analysis of brain MDA and GSH level after SQ22536 administration. Data are presented as mean ​± ​SD. ∗p ​< ​0.05 vs. sham group; @p ​< ​0.05 vs. ICH ​+ ​vehicle group; &p ​< ​0.05 vs. ICH ​+ ​PAL ​+ ​DMSO group. (n ​= ​6/group). #p ​< ​0.05 vs ICH ​+ ​Vehicle ​+ ​Scr siRNA group; $p ​< ​0.05 vs ICH ​+ ​PAL ​+ ​Scr siRNA group; ∗p ​< ​0.05 vs. sham group; @p ​< ​0.05 vs. ICH ​+ ​vehicle group; &p ​< ​0.05 vs. ICH ​+ ​PAL ​+ ​DMSO group. (n ​= ​6/group).
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Neuroprotection by paricalcitol in vivo is VDR-dependent and mediated by the cAMP-PKA-DRP1 pathway. (A, B) Representative Western blot bands and quantitative analysis for VDR, ACSL4, and 4HNE after VDR siRNA administration in each group. (C) Results of the modified Garcia test, forelimb placement test, and corner turn test at 24 ​h post-ICH after VDR siRNA administration (n ​= ​6/group). (D) Representative Western blot bands for cAMP, p-PKA, PKA, p-DRP1 (Ser637), ACSL4, and 4HNE at 24 ​h post-ICH with SQ22536 injection. (E) Quantitative analysis of cAMP, p-PKA, PKA, p-DRP1 (Ser637), ACSL4, and 4HNE levels. (F) Results of the modified Garcia test, forelimb placement test, and corner turn test after SQ22536 administration. (G, H) Quantitative ELISA analysis of brain MDA and GSH level after SQ22536 administration. Data are presented as mean ​± ​SD. ∗p ​< ​0.05 vs. sham group; @p ​< ​0.05 vs. ICH ​+ ​vehicle group; &p ​< ​0.05 vs. ICH ​+ ​PAL ​+ ​DMSO group. (n ​= ​6/group). #p ​< ​0.05 vs ICH ​+ ​Vehicle ​+ ​Scr siRNA group; $p ​< ​0.05 vs ICH ​+ ​PAL ​+ ​Scr siRNA group; ∗p ​< ​0.05 vs. sham group; @p ​< ​0.05 vs. ICH ​+ ​vehicle group; &p ​< ​0.05 vs. ICH ​+ ​PAL ​+ ​DMSO group. (n ​= ​6/group).

Journal: Neurotherapeutics

Article Title: Paricalcitol-mediated vitamin D receptor activation attenuates neuronal ferroptosis via cAMP-PKA-DRP1 signaling pathway after intracerebral hemorrhage

doi: 10.1016/j.neurot.2025.e00767

Figure Lengend Snippet: Neuroprotection by paricalcitol in vivo is VDR-dependent and mediated by the cAMP-PKA-DRP1 pathway. (A, B) Representative Western blot bands and quantitative analysis for VDR, ACSL4, and 4HNE after VDR siRNA administration in each group. (C) Results of the modified Garcia test, forelimb placement test, and corner turn test at 24 ​h post-ICH after VDR siRNA administration (n ​= ​6/group). (D) Representative Western blot bands for cAMP, p-PKA, PKA, p-DRP1 (Ser637), ACSL4, and 4HNE at 24 ​h post-ICH with SQ22536 injection. (E) Quantitative analysis of cAMP, p-PKA, PKA, p-DRP1 (Ser637), ACSL4, and 4HNE levels. (F) Results of the modified Garcia test, forelimb placement test, and corner turn test after SQ22536 administration. (G, H) Quantitative ELISA analysis of brain MDA and GSH level after SQ22536 administration. Data are presented as mean ​± ​SD. ∗p ​< ​0.05 vs. sham group; @p ​< ​0.05 vs. ICH ​+ ​vehicle group; &p ​< ​0.05 vs. ICH ​+ ​PAL ​+ ​DMSO group. (n ​= ​6/group). #p ​< ​0.05 vs ICH ​+ ​Vehicle ​+ ​Scr siRNA group; $p ​< ​0.05 vs ICH ​+ ​PAL ​+ ​Scr siRNA group; ∗p ​< ​0.05 vs. sham group; @p ​< ​0.05 vs. ICH ​+ ​vehicle group; &p ​< ​0.05 vs. ICH ​+ ​PAL ​+ ​DMSO group. (n ​= ​6/group).

Article Snippet: To investigate the role of the cAMP-PKA-DRP1 signaling pathway, the cAMP inhibitor SQ22536 (Tocris Bioscience, USA, Cat#1435/10) was administered intracerebroventricularly (ICV) at a dose of 2 nmol per mouse [ ].

Techniques: In Vivo, Western Blot, Modification, Injection, Enzyme-linked Immunosorbent Assay

Paricalcitol protects neurons from hemin-induced death in vitro via a VDR- and cAMP-dependent mechanism. (A) Flow cytometry dot plots and quantification of neuronal survival and apoptosis. PI (−)/Annexin V (+) indicates early apoptotic neurons. (B) Flow cytometry dot plots and quantification of ROS levels after hemin stimulation. (C) Flow cytometry dot plots and quantification of JC-1 staining for mitochondrial membrane potential. (D) Representative immunofluorescence images of JC-1 staining after hemin stimulation. (E) Flow cytometry dot plots and quantification of neuronal survival and apoptosis after VDR knockdown. PI (−)/Annexin V (+) indicates early apoptotic neurons. (F) Flow cytometry dot plots and quantification of neuronal survival and apoptosis after SQ22536 administration. PI (−)/Annexin V (+) indicates early apoptotic neurons. %p ​< ​0.05 vs. Hemin ​+ ​Scr siRNA group. ¥p ​< ​0.05 vs. Hemin ​+ ​PAL ​+ ​DMSO group.

Journal: Neurotherapeutics

Article Title: Paricalcitol-mediated vitamin D receptor activation attenuates neuronal ferroptosis via cAMP-PKA-DRP1 signaling pathway after intracerebral hemorrhage

doi: 10.1016/j.neurot.2025.e00767

Figure Lengend Snippet: Paricalcitol protects neurons from hemin-induced death in vitro via a VDR- and cAMP-dependent mechanism. (A) Flow cytometry dot plots and quantification of neuronal survival and apoptosis. PI (−)/Annexin V (+) indicates early apoptotic neurons. (B) Flow cytometry dot plots and quantification of ROS levels after hemin stimulation. (C) Flow cytometry dot plots and quantification of JC-1 staining for mitochondrial membrane potential. (D) Representative immunofluorescence images of JC-1 staining after hemin stimulation. (E) Flow cytometry dot plots and quantification of neuronal survival and apoptosis after VDR knockdown. PI (−)/Annexin V (+) indicates early apoptotic neurons. (F) Flow cytometry dot plots and quantification of neuronal survival and apoptosis after SQ22536 administration. PI (−)/Annexin V (+) indicates early apoptotic neurons. %p ​< ​0.05 vs. Hemin ​+ ​Scr siRNA group. ¥p ​< ​0.05 vs. Hemin ​+ ​PAL ​+ ​DMSO group.

Article Snippet: To investigate the role of the cAMP-PKA-DRP1 signaling pathway, the cAMP inhibitor SQ22536 (Tocris Bioscience, USA, Cat#1435/10) was administered intracerebroventricularly (ICV) at a dose of 2 nmol per mouse [ ].

Techniques: In Vitro, Flow Cytometry, Staining, Membrane, Immunofluorescence, Knockdown