pp1 γ  (Santa Cruz Biotechnology)

Journal: Cellular & Molecular Biology Letters

Article Title: HNF4A P2 isoform alleviates kidney fibrosis by inhibiting dedifferentiation of proximal tubular cells through JAG1/NOTCH signaling

doi: 10.1186/s11658-025-00845-0

Figure Lengend Snippet: p-SRC selectively inhibits the function of HNF4A P1 instead of HNF4A P2. A Western blots of p-SRC/SRC in HK-2 cells treated with different concentration of TGF-β1. B , C Representative Western blots and the relative quantitation of protein expression of p-SRC of kidney tissues from UIRI and UUO ( n = 6). D , E Representative Western blots and the relative quantitation of protein expression of p-SRC in HK-2 cells treated with different concentration of TGF-β1, with or without HNF4A P1/P2 overexpression ( n = 3). F Phosphorylation of HNF4A P1/P2 protein in HK-2 cells treated with different concentration of TGF-β1 with or without HNF4A P1/P2 plasmids were determined via IP and Western blot assays ( n = 3). G – M Representative Western blots and the relative quantitation of protein expression of SNAI1, VIM, JAG1, NOTCH1, NOTCH2, and NOTCH3 of HK-2 cells treated with different concentration of TGF-β1, with or without HNF4A P1 overexpression and PP1 ( n = 3). N , O The mRNA level of SNAI1 and VIM in HK-2 cells treated with different concentration of TGF-β1, with or without HNF4A P1 overexpression and PP1. P , Q Representative Western blots and the relative quantitation of protein expression of α-SMA of NRK-49F cells treated with different conditioned medium derived from HK2 cells ( n = 3). R qPCR analysis of ChIP assays with anti-HNF4A P1 antibody on HK-2 cells from different groups and relative quantitation ( n = 3). Data are presented as mean ± SD; ANOVA test was used; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: To induce fibroblast activation, following overnight starvation in serum-free medium, subconfluent NRK-49F fibroblasts were incubated with tubular cell-conditioned medium for 48 h. To inhibit SRC activation, HK-2 and TCMK-1 cells were incubated with 10 ng/ml TGF-β1 for 48 h in the presence of PP1 (5 μM, TargetMol, MA, China).

Techniques: Western Blot, Concentration Assay, Quantitation Assay, Expressing, Over Expression, Phospho-proteomics, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Fluoxetine Repurposing Mitigates Alzheimer’s Disease Pathology via the GSK3β–CREB–ADAM10 Axis

doi: 10.3390/ijms27062676

Figure Lengend Snippet: Fluoxetine modulates CaMKII-GSK3β-CREB signaling and ADAM10 expression in vitro. ( A ) Schematic diagram of the experimental design. SH-SY5Y cells were seeded and pretreated with fluoxetine (10 μM) for 12 h, followed by treatment with the CaMKII inhibitor 1-NA-PP1 (10 or 20 μM) or the GSK3β inhibitor IX (5 or 10 μM) for 1 h prior to cell harvest. ( B ) Representative Western blot analysis for phosphorylated and total forms of CaMKII (Thr286), GSK3β (Ser9), and CREB (Ser133). ( C ) Quantification of relative phosphorylation levels normalized to the corresponding total protein. ( D ) Representative Western blot analysis of signaling and APP-processing-related proteins following fluoxetine and/or GSK3β inhibitor treatment. ( E ) Quantification of protein expression levels normalized to β-actin (for protein) or to the corresponding total protein (for phosphorylated forms), as indicated. Data are presented as mean ± SEM ( n = 3 independent experiments). Individual data points represent independent biological replicates. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. * p < 0.05, ** p < 0.01 vs. control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. fluoxetine alone; ns, not significant. Created in BioRender. Son, Y. (2026) https://BioRender.com/s91n646 (accessed on 31 January 2026).

Article Snippet: After 24 h, 10 μM fluoxetine was added to the culture dishes for 12 h. 1-Naphthyl PP1 (1-NA-PP1, HY-13941), a CAMKII inhibitor, and GSK3 inhibitor IX (HY-10580), a GSK3β inhibitor, were purchased from MedChem Express (Princeton, NJ, USA).

Techniques: Expressing, In Vitro, Western Blot, Phospho-proteomics, Control