Journal: Scientific Reports

Article Title: Podocyte injury in diabetic nephropathy: implications of angiotensin II – dependent activation of TRPC channels

doi: 10.1038/srep17637

Figure Lengend Snippet: ( a ) A representative Western blot and a summary graph for the densitometry values of the TRPC6 channel signal blotted from the cortical kidney lysates of the control and STZ-treated rats (terminal point, 11 weeks after STZ injection). Loading control for the representative blot (β-actin) and number of animals per group analyzed in the summary graph (N) are shown. Asterisk denotes statistical significance (P < 0.05). ( b ) Images of the representative immunohistochemical stainings for TRPC6 in the control and STZ-treated rats. Magnification – 20 x; scale bar is shown.

Article Snippet: Western blot analysis of the blood-free kidney cortex was performed as reported previously using antibodies against an intracellular epitope of the TRPC6 ion channel (cat no ACC-017, Alomone Labs).

Techniques: Western Blot, Control, Injection, Immunohistochemical staining, Immunohistochemistry

Journal: Clinical and Translational Medicine

Article Title: Low‐glucose‐sensitive TRPC6 dysfunction drives hypoglycemia‐induced cognitive impairment in diabetes

doi: 10.1002/ctm2.205

Figure Lengend Snippet: RH specifically represses hippocampal TRPC6 channel in diabetic mice. (A) Relative TRPC6 mRNA expression (n = 6) in PC12 cells cultured in medium with normal glucose concentration (NG, 5.6 mM) or medium with high glucose concentration (HG, 25 mM). NG+LG, cells cultured in NG medium and received three times of low glucose stimulation (12 hours per times, LG, 3.0 mM); HG+LG, cells cultured in HG medium and received three times of low glucose stimulation (12 hours per times, LG, 3.0 mM). (B) Representative Western blots of TRPC6 protein in PC12 cells in four groups. GAPDH served as a loading control. Quantitative data were shown on the right (n = 4). (C) Relative mRNA expression levels of TRPC6 in hippocampus from nondiabetic (ND) mice, ND mice with recurrent moderate hypoglycemia (ND‐RH), mice with STZ‐induced type 1 diabetes mellitus (DM), and DM mice with RH (DM‐RH) (n = 6 samples from six mice). (D) Representative Western blots of TRPC6 in hippocampus. GAPDH served as a loading control. Quantitative results are shown on the right (n = 3 tissues from three mice). TPRC6 −/− , TRPC6 global knockout mice. (E) Relative mRNA expression levels of TRPC1, TRPC3, TRPC4, TRPC5, and TRPC7 in hippocampus (n = 6–8 samples from four mice). * P < .05, ** P < .01, *** P < .001. Statistical significance was assessed using a one‐way ANOVA or Kruskal‐Wallis test

Article Snippet: The primary antibodies were used, including anti‐TRPC1 (ACC‐010, 120 kDa), anti‐TRPC3 (ACC‐016, 100 kDa), anti‐TRPC4 (ACC‐018, 97 kDa), anti‐TRPC5 (ACC‐020, 100 kDa), anti‐TRPC6 antibody (ACC‐017, 110 kDa), anti‐TRPC7 antibody (ACC‐066, 95 kDa), anti‐TRPV1 (ACC‐030, 90 kDa), and anti‐TRPV4 (ACC‐034, 100 kDa) from Alomone; anti‐LKB1 (PA5‐96062, 48 kDa), anti‐phospho‐CaMMK2 (PA5‐105225, 65 kDa), and anti‐phospho‐DRP1 (S616, PA5‐64821, predicted to react with Ser585 in rats and Ser600 in mice, 83 kDa) antibodies from Invitrogen; anti‐CaMKK2 (ab135979, 65 kDa), anti‐Drp1 (ab184247, 83 kDa), anti‐phospho‐DRP1 (S637, ab193216, predicted to react with Ser656 in rats and Ser616 in mice, 83 kDa), anti‐PSD95 (ab238135, 80 kDa), anti‐Synapsin I (ab254349, 74 kDa), and anti‐SYP (ab32127, 34 kDa) antibodies from Abcam; anti‐AMPKα (2532S, 62 kDa) and anti‐phospho‐AMPKα (Thr172, 2535S, 62 kDa) antibodies from CST, followed by incubation with the secondary antibodies (ZSGB‐BIO).

Techniques: Expressing, Cell Culture, Concentration Assay, Western Blot, Knock-Out

Journal: Clinical and Translational Medicine

Article Title: Low‐glucose‐sensitive TRPC6 dysfunction drives hypoglycemia‐induced cognitive impairment in diabetes

doi: 10.1002/ctm2.205

Figure Lengend Snippet: RH specifically reduces TRPC6 expression in the white blood cells from diabetic patients. (A) Relative mRNA expression of TRPC6 in human blood white blood cells from normal subject (n = 14 samples from 14 patients), type 2 diabetic patients without hypoglycemia episodes (T2DM, n = 18 samples from 18 patients) or diabetic patients with RH (T2DM‐RH, n = 8 samples from eight patients). (B‐E) Relative mRNA expression of TRPC1, TRPC3, TRPC5, and TRPC7 in human white blood cells. (F‐I) Representative image showed Ca 2+ influx in white blood cells from a normal subject (F), a T2DM patient without RH (G), and a T2DM patient with RH and quantitative data are shown on the right (I). Ca 2+ (1 mM, Cacl 2 ) and/or hyperforin (Hyp, 10 µM) was added at the point 25S; * P < 0.01, ** P < 0.01, and *** P < 0.001. Statistical significance was assessed using Kruskal‐Wallis test followed by Dunn's multiple comparisons test

Article Snippet: The primary antibodies were used, including anti‐TRPC1 (ACC‐010, 120 kDa), anti‐TRPC3 (ACC‐016, 100 kDa), anti‐TRPC4 (ACC‐018, 97 kDa), anti‐TRPC5 (ACC‐020, 100 kDa), anti‐TRPC6 antibody (ACC‐017, 110 kDa), anti‐TRPC7 antibody (ACC‐066, 95 kDa), anti‐TRPV1 (ACC‐030, 90 kDa), and anti‐TRPV4 (ACC‐034, 100 kDa) from Alomone; anti‐LKB1 (PA5‐96062, 48 kDa), anti‐phospho‐CaMMK2 (PA5‐105225, 65 kDa), and anti‐phospho‐DRP1 (S616, PA5‐64821, predicted to react with Ser585 in rats and Ser600 in mice, 83 kDa) antibodies from Invitrogen; anti‐CaMKK2 (ab135979, 65 kDa), anti‐Drp1 (ab184247, 83 kDa), anti‐phospho‐DRP1 (S637, ab193216, predicted to react with Ser656 in rats and Ser616 in mice, 83 kDa), anti‐PSD95 (ab238135, 80 kDa), anti‐Synapsin I (ab254349, 74 kDa), and anti‐SYP (ab32127, 34 kDa) antibodies from Abcam; anti‐AMPKα (2532S, 62 kDa) and anti‐phospho‐AMPKα (Thr172, 2535S, 62 kDa) antibodies from CST, followed by incubation with the secondary antibodies (ZSGB‐BIO).

Techniques: Expressing