pik3ip1  (Genechem)

Journal: Experimental and Therapeutic Medicine

Article Title: CDX2-UPK1B-PIK3IP1-PI3K/AKT signaling axis regulates gastric cancer cell invasion and migration and influences patient prognosis

doi: 10.3892/etm.2026.13179

Figure Lengend Snippet: UPK1B activates PI3K/AKT signaling by antagonizing the inhibitory regulator PIK3IP1 in gastric cancer cells. (A) Venn diagram showing that PIK3IP1 was identified as a putative UPK1B-interacting partner based on BioGRID and HIPPIE protein-protein interaction databases. (B) UPK1B and PIK3IP1 co-localized in the cytoplasm and plasma membrane of MKN45 cells. (C) Interaction between UPK1B and PIK3IP1 in MKN45 cells. (D) Knockdown of PIK3IP1 in MKN45 cells. (E) Silencing PIK3IP1 in UPK1B-knockdown MKN45 cells restored PI3K/AKT pathway activation. Knockdown of PIK3IP1 reversed the decrease in (F) migration/invasion and (G) wound-healing capacity in UPK1B-silenced MKN45 cells. UPK1B, uroplakin 1B; p-, phosphorylated; si, small interfering RNA; sh, short hairpin RNA; NC, negative control; PIK3IP1, PI3K inhibitor interacting protein 1; HIPPIE, Human Integrated Protein-Protein Interaction Reference; IP, immunoprecipitation.

Article Snippet: Cells were also transfected with small interfering RNAs (siRNAs) targeting CDX2 or PIK3IP1 , with a universal non-targeting scrambled siRNA (si-NC) as the negative control , obtained from GeneChem, Inc. For UPK1B and CDX2 overexpression, the p-TSB-CMV-UPK1B and p-TSB-CMV-CDX2 expression vectors [Shanghai Genomeditech Co., Ltd.] and the corresponding empty p-TSB-CMV vector (negative control) were used.

Techniques: Clinical Proteomics, Membrane, Knockdown, Activation Assay, Migration, Small Interfering RNA, shRNA, Negative Control, Immunoprecipitation

Journal: bioRxiv

Article Title: Downregulation of PIK3IP1/TrIP on T cells is controlled by TCR signal strength, PKC, and metalloprotease-mediated cleavage

doi: 10.1101/2024.04.29.591680

Figure Lengend Snippet: A , ExCYT clustering analysis of flow cytometry data identifies major splenic populations including B cells (CD19 + ), CD4 (CD3 + CD4 + FoxP3 - ) and CD8 (CD3 + CD8 + ) T cells, regulatory T cells (CD3 + CD4 + FoxP3 + ), DCs (CD11c + ), macrophages (CD11b + F4/80 + ), and neutrophils (CD11b + Gr-1 Hi ) . B , Heatmap of TrIP protein expression overlayed across the splenic populations. C, Quantification of TrIP MFI across each of the immune cell populations. D, Histograms depicting the range of TrIP expression in the indicated populations. E, Staining of WT vs TrIP knockout ( Pik3ip1 fl/fl E8i cre ) CD8 + T cells (CD3 + CD8 + ) from naïve spleens. F, Gating strategy for naïve (CD62L + ) and antigen-experienced (CD44 + ) CD8 + T cells and corresponding TrIP expression. G, Quantification of TrIP-AF647 MFI in the CD8 + T cell subpopulations, stratified by CD44 vs CD62L expression. H, Stimulation of whole WT P14 TCR Tg splenocytes with 200 ng/ml of WT gp33 peptide. TrIP and TCR Vα2 expression were followed over 24 hrs by flow cytometry. I, Quantification of TrIP expression MFI on CD8+ T cells in the 24 hrs following activation in vitro .

Article Snippet: For the initial antibody validation experiments, full-length human PIK3IP1 and full-length mouse Pik3ip1 gBlocks (IDT) were cloned into the pcDNA3.1 expression vector via restriction enzyme cloning.

Techniques: Flow Cytometry, Expressing, Staining, Knock-Out, Activation Assay, In Vitro

Journal: bioRxiv

Article Title: Downregulation of PIK3IP1/TrIP on T cells is controlled by TCR signal strength, PKC, and metalloprotease-mediated cleavage

doi: 10.1101/2024.04.29.591680

Figure Lengend Snippet: A , ExCYT clustering analysis of flow cytometry data identifies major splenic populations including B cells (CD19 + ), CD4 (CD3 + CD4 + FoxP3 - ) and CD8 (CD3 + CD8 + ) T cells, regulatory T cells (CD3 + CD4 + FoxP3 + ), DCs (CD11c + ), macrophages (CD11b + F4/80 + ), and neutrophils (CD11b + Gr-1 Hi ) . B , Heatmap of TrIP protein expression overlayed across the splenic populations. C, Quantification of TrIP MFI across each of the immune cell populations. D, Histograms depicting the range of TrIP expression in the indicated populations. E, Staining of WT vs TrIP knockout ( Pik3ip1 fl/fl E8i cre ) CD8 + T cells (CD3 + CD8 + ) from naïve spleens. F, Gating strategy for naïve (CD62L + ) and antigen-experienced (CD44 + ) CD8 + T cells and corresponding TrIP expression. G, Quantification of TrIP-AF647 MFI in the CD8 + T cell subpopulations, stratified by CD44 vs CD62L expression. H, Stimulation of whole WT P14 TCR Tg splenocytes with 200 ng/ml of WT gp33 peptide. TrIP and TCR Vα2 expression were followed over 24 hrs by flow cytometry. I, Quantification of TrIP expression MFI on CD8+ T cells in the 24 hrs following activation in vitro .

Article Snippet: For the initial antibody validation experiments, full-length human PIK3IP1 and full-length mouse Pik3ip1 gBlocks (IDT) were cloned into the pcDNA3.1 expression vector via restriction enzyme cloning.

Techniques: Flow Cytometry, Expressing, Staining, Knock-Out, Activation Assay, In Vitro

Journal: Genome Medicine

Article Title: Acute ischemia induces spatially and transcriptionally distinct microglial subclusters

doi: 10.1186/s13073-023-01257-5

Figure Lengend Snippet: Glucocorticoids triggers IPAM formation. A Upstream IPA of top 200 differentially expressed genes in IPAM (Fisher’s exact test, Benjamini–Hochberg FDR). B Histograms visualizing targeted metabolomics results in the sham, infarct, and peri-infarct group brain samples from sham and MCAO-12 h mice. n = 6/group. C – D Heatmap showing the expression of several ICAM-specific or IPAM-specific marker genes upon DEX (5 nM, 24 h) ( C ) or CORT (1 μM, 24 h) ( D ) stimulation. n = 3/group. E – F TTC staining of brains from CON, CORT, and RU486 group ( E ). The infarct volume was quantified ( F ). n = 6/group. G – H mNSS were performed at 1 day ( G ) and 3 days ( H ) after MCAO to evaluate the neurological deficits of each group. n = 7–14/group. I Grip strength was performed at 1 day and 3 days after MCAO to evaluate the neurological deficits of each group. n = 7–14/group. J Representative TUNEL images co-stained with neuronal-marker NEUN within the ischemic regions in the MCAO 1d brains from CON, CORT, and RU486 groups (scale bar: 50 µm). K Proportion of TUNEL + /NeuN + cells in NeuN + cells was quantified. n = 3/group. L Expression of PIK3IP1 in TMEM119. + cells was quantified. The fluorescence intensity of PIK3IP1 in the CORT and RU486 groups was normalized to the mean value of that measured in the CON group. n = 3/group. M Representative immunostaining of IPAM-specific marker PIK3IP1 and microglia-specific marker TMEM119 in the peri-infarct within MCAO-1d sections from CON, CORT, and RU486 groups (scale bar: 20 µm). In B , F – H , K , and L , one-way ANOVA with Tukey’s post hoc test. In I , two-way ANOVA with Tukey’s post hoc test. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies were directed against Tmem119 (1:500, Synaptic Systems, Germany, 400 011), Tmem119 (1:100, Abcam, USA, ab209064), LGALS3 (1:500, Biolegend, USA, 125,401), PIK3IP1 (1:50, Santa Cruz Biotechnology, USA, sc-365777), HMGB1 (1:500, Abcam, USA, ab79823), and NEUN (1:500, Abcam, USA, ab104224).

Techniques: Expressing, Marker, Staining, TUNEL Assay, Fluorescence, Immunostaining