trpc1  (Proteintech)

Journal: iScience

Article Title: The role of arachidonic acid metabolites in the subtype classification and pathogenesis of primary aldosteronism

doi: 10.1016/j.isci.2025.114598

Figure Lengend Snippet: Arachidonic acid promotes Ca 2+ uptake and increases CYP11B2 expression in adrenal cortical cells (A) Confirmation of the expression of CYP11B2 in primary adrenal cortical cells. Scale bars, 50 μm. (B) Cell viability of primary adrenal cortical cells treated with different doses of arachidonic acid (AA) ( n = 4). (C) Aldosterone levels in cellular supernatant of primary adrenal cortical cells treated with different doses of AA ( n = 6). (D) Aldosterone levels in cellular supernatant of primary adrenal cortical cells treated with different doses of Ang II (left) or endothelin-1 (right) along with AA ( n = 3). (E) Representative western blots showing levels of CYP11B2, CYP11B1, CYP17A1, and HSD3B2 in primary adrenal cortical cells treated with vehicle or AA. The quantitative results are shown on the right ( n = 3). (F) Changes of cytoplasmic Ca 2+ , labeled with Fura-2 AM, in primary adrenal cortical cells treated with 1 mM thapsigargin (TG) stimulation in a 1 mM extracellular Ca 2+ solution after preincubation with vehicle or AA ( n = 12). (G) Changes of mitochondrial Ca 2+ , labeled with Rhod-2 AM, in digitonin-permeabilized primary adrenal cortical cells treated with 200 μM ATP stimulation in a 300 μM extracellular Ca 2+ solution after preincubation with vehicle or AA ( n = 12). (H) Changes of endoplasmic reticulum (ER) Ca 2+ , labeled with Mag Fura-2 AM, in digitonin-permeabilized primary adrenal cortical cells treated with ATP stimulation in a Ca 2+ -free extracellular solution after preincubation with vehicle or AA ( n = 12). (I) Representative western blots showing levels of KCNJ5, Na + /K + ATPase alpha-1 subunit, NCX-1, Letm 1, MCU, VDAC, RyR2, and IP 3 R in primary adrenal cortical cells treated with vehicle or AA. (Left) Representative western blots showing levels of TRPC1, TRPC3, TRPC6, TRPV1, and TRPV4 in primary adrenal cortical cells treated with vehicle or AA. (Right) The quantitative results are shown on the left ( n = 3). The results are expressed as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001 compared with vehicle group; # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 compared with 1 μM AA group by one-way ANOVA (B and C) and by Student’s t test (D–I).

Article Snippet: TRPC1 , Alomone , Cat#ACC-010; RRID: AB_2040234.

Techniques: Expressing, Western Blot, Labeling

Journal: Cell Death Discovery

Article Title: Lysine attenuates acute lung injury by restoring α-tubulin acetylation and ciliary activity

doi: 10.1038/s41420-026-03025-x

Figure Lengend Snippet: Calcium image analysis in PQ-injured A549 cells compared to normal cells ( A ) and PQ-injured A549 cells w/wo lysine supplementation ( B ) with thapsigargin (TG) or ionomycin (IONO) stimulation. Mean ± SEM. C NFAT luciferase expression in A549 cells treated w/wo 800 μM PQ, together with 0-, 1-, 2.5-, or 5-fold lysine as in the culture medium as indicated, for 24 h. Mean ± SD, *** P < 0.001; Two-Way ANOVA. n = 3. NS not significant. D WB analysis of E-Cadherin, ZO-1, EPCAM and acetyl-α-Tubulin in PQ-injured A549 cells treated w/wo 5 μM SKF-96365 (SKF). E Co-IP analysis of STIM1 association with ORAI1 or TRPC1 in PQ-injured A549 cells with indicated concentration, together w/wo 5 mM lysine. F Immunofluorescence analysis of Myc and ARL13B in PQ-injured TRPC1-Myc stably expressed A549 cells w/wo lysine supplementation. Scale bar, 10 μm.

Article Snippet: Specific antibodies were used for STIM1 (Cell Signaling Technology, 5668, 1:3000), ORAI1 (Santa Cruz, sc-377281, 1:500), and TRPC1 (Proteintech, 19482, 1:2000).

Techniques: Luciferase, Expressing, Co-Immunoprecipitation Assay, Concentration Assay, Immunofluorescence, Stable Transfection

Journal: Cell Death Discovery

Article Title: Lysine attenuates acute lung injury by restoring α-tubulin acetylation and ciliary activity

doi: 10.1038/s41420-026-03025-x

Figure Lengend Snippet: A GSEA analysis uncovered top-ranked molecular functions enhanced in PQ-injured lung tissues with lysine supplementation compared to the untreated group. B KEGG enrichment of the top 20 upregulated pathways in PQ-injured lung tissues with lysine supplementation compared to the untreated group. C Quantification of NADP and NADPH (left) or NAD and NADH (right) in PQ-injured A549 cells w/wo lysine supplementation. D ELISA analysis of acetyl coenzyme A (Acetyl-CoA) in PQ-injured A549 cells w/wo lysine supplementation. E WB analysis of E-Cadherin and acetyl-α-Tubulin in PQ-injured A549 cells treated w/wo 1 mM sodium acetate (SA). F WB analysis of ZO-1, E-Cadherin, acetyl-α-Tubulin and EPCAM in PQ-injured A549 cells w/wo lysine supplementation, together w/wo 0.5 μM GM-90257 (GM). G Calcium image analysis in PQ-injured A549 cells w/wo lysine supplementation, together w/wo 0.5 μM GM-90257 treatment. The cells were transiently stimulated with thapsigargin (TG) or ionomycin (IONO). Mean ± SEM. H Co-IP analysis of STIM1 association with TRPC1 in PQ-injured A549 cells w/wo lysine supplementation, together w/wo GM-90257 treatment. Immunofluorescence analysis of ARL13B, SFTPC, and HOPX in A549 cells ( I ) or normal lung tissues ( J ). Scale bar, 10 μm. Mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001; Two-Way ANOVA. NS not significant.

Article Snippet: Specific antibodies were used for STIM1 (Cell Signaling Technology, 5668, 1:3000), ORAI1 (Santa Cruz, sc-377281, 1:500), and TRPC1 (Proteintech, 19482, 1:2000).

Techniques: Enzyme-linked Immunosorbent Assay, Co-Immunoprecipitation Assay, Immunofluorescence

Journal: Biological Research

Article Title: Shear stress-induced Ca 2+ influx triggers endoplasmic reticulum stress and cardiomyocyte apoptosis: implications for mitral regulation

doi: 10.1186/s40659-026-00671-4

Figure Lengend Snippet: The effects of TRPC1 knockdown on real-time Ca 2+ measurements. A To optimize the TRPC1 knockdown efficiency, HL-1 cells were transfected with siCtrl, siTRPC1#1, or siTRPC1#2 for 24 and 48 h. Parental cells were not transfected with siRNA. B Images were taken at 1 frame/3 s to record Ca 2+ activity in 2 mM Ca 2+ buffer. HL-1 cells were pretreated with fura-2 (2 μM) for 30 min. The panels are the Ca 2+ curve of the conditions of siCtrl, siCtrl with shear stress, siTRPC1 and siTRPC1 with shear stress. C The columns represent the amplitude of the Ca 2+ oscillations and D the changes in the Ca 2+ level in each group. All columns are presented as the mean ± SEM of at least three independent experiments with at least 20 cells of each group. The results were analyzed via one-way ANOVA. *** p < 0.001

Article Snippet: The samples were analyzed using antibodies against Grp78 (BD, 610978), ATF6 (Santa Cruz, sc166659), p-PERK (Cell Signaling, 3179), PERK (Cell Signaling, 3192), p-IRE1α (Abcam, ab48187), IRE1α (Abcam, ab37073), p-eIF2α (Cell Signaling, 9721), eIF2α (Cell Signaling, 9722), ATF4 (Santa Cruz, sc390063), CHOP (Cell Signaling, 2895), Bax (Genetex, GTX109683), Bcl-2 (BD, 610538), Bak (Cell Signaling, 12105), caspase 9 (Santa Cruz, sc56076), caspase 12 (Cell Signaling, 35965), caspase 3 (Abcam, ab32042), caspase 8 (Proteintech, 13423-1-AP), Bid (Novus, 10988-1-AP), and transient receptor potential canonical 1 (TRPC1) (Santa Cruz, sc133076).

Techniques: Knockdown, Transfection, Activity Assay, Shear