rabbit anti ucp1  (Jackson Immuno)

 
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    Name:
    Streptavidin
    Description:
    Steptavidin a bacterial protein isolated from Streptomyces avidinii is similar to egg white avidin in its ability to bind biotin and has been used as a replacement for egg white avidin because of its more favorable chemical properties Conjugates of streptavidin are recommended for use with Biotin SP conjugated antibodies and Biotin SP conjugated ChromPure proteins
    Catalog Number:
    016-000-084
    Price:
    63
    Conjugate:
    Unconjugated
    Size:
    mg
    Category:
    Streptavidin
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    Structured Review

    Jackson Immuno rabbit anti ucp1
    Streptavidin
    Steptavidin a bacterial protein isolated from Streptomyces avidinii is similar to egg white avidin in its ability to bind biotin and has been used as a replacement for egg white avidin because of its more favorable chemical properties Conjugates of streptavidin are recommended for use with Biotin SP conjugated antibodies and Biotin SP conjugated ChromPure proteins
    https://www.bioz.com/result/rabbit anti ucp1/product/Jackson Immuno
    Average 94 stars, based on 1849 article reviews
    Price from $9.99 to $1999.99
    rabbit anti ucp1 - by Bioz Stars, 2020-09
    94/100 stars

    Images

    1) Product Images from "α-ketoglutaric acid stimulates muscle hypertrophy and fat loss through OXGR1-dependent adrenal activation"

    Article Title: α-ketoglutaric acid stimulates muscle hypertrophy and fat loss through OXGR1-dependent adrenal activation

    Journal: bioRxiv

    doi: 10.1101/796037

    Adrenal specific overexpression of OXGR1 enhances stimulatory effects of AKG on thermogenesis and lipolysis (A). The validation of OXGR1 overexpression. The mRNA expression of OXGR1 was determined in the adrenal glands from male WT control, WT injected with HBAAV2/9-GFP, and WT injected with HBAAV2/9-OXGR1 (OXGR1OE AG ) mice (n=5 per group). (B-C). Body weight gain (B) and cumulative food intake (C) of OXGR1OE AG . Male C57BL/6 mice (8 weeks) were adrenal-specifically injected with control HBAAV2/9-GFP or HBAAV2/9-OXGR1. Two weeks after injections, mice were switched to HFD and further divided into two groups, receiving tap water or water supplemented with 2% AKG for 12 weeks (n = 8 per group). (D-E). Representative image of body composition (D) and fat and lean mass index (E) of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 8 per group). (F-G). Weight index of gWAT (F) and iWAT (G) in male OXGR1OE AG mice treated with AKG for 12 weeks (n = 6 per group). (H-I). Immunoblots (H) and quantification (I) of p-HSL and ATGL protein in the gWAT of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 4 per group). (J). Immunoblots and quantification of UCP1 protein in the BAT of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 4 per group). (K) Serum E level in male OXGR1OE AG mice treated with AKG for 12 weeks (n= 8 per group). (L-O). Oxygen consumption (L-M) and RER (N-O) of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 8 per group). (P-Q). Representative images (P) and quantification (Q) of gWAT and iWAT HE staining from male OXGR1OE AG mice treated with AKG for 12 weeks (n = 6 per group). (R-S). Representative images (R) and quantification (S) of p-HSL DAB staining from male OXGR1OE AG mice treated with AKG for 12 weeks (n = 6 per group). Results are presented as mean ± SEM. In (A), ** p≤0.01 by non-paired Student’s t test. In (B-C), *p≤0.05, **p≤0.01 by two-way ANOVA followed by post hoc Bonferroni tests. In (E-G), (I-K), (M), (O), (Q) and (S), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Turkey’s tests.
    Figure Legend Snippet: Adrenal specific overexpression of OXGR1 enhances stimulatory effects of AKG on thermogenesis and lipolysis (A). The validation of OXGR1 overexpression. The mRNA expression of OXGR1 was determined in the adrenal glands from male WT control, WT injected with HBAAV2/9-GFP, and WT injected with HBAAV2/9-OXGR1 (OXGR1OE AG ) mice (n=5 per group). (B-C). Body weight gain (B) and cumulative food intake (C) of OXGR1OE AG . Male C57BL/6 mice (8 weeks) were adrenal-specifically injected with control HBAAV2/9-GFP or HBAAV2/9-OXGR1. Two weeks after injections, mice were switched to HFD and further divided into two groups, receiving tap water or water supplemented with 2% AKG for 12 weeks (n = 8 per group). (D-E). Representative image of body composition (D) and fat and lean mass index (E) of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 8 per group). (F-G). Weight index of gWAT (F) and iWAT (G) in male OXGR1OE AG mice treated with AKG for 12 weeks (n = 6 per group). (H-I). Immunoblots (H) and quantification (I) of p-HSL and ATGL protein in the gWAT of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 4 per group). (J). Immunoblots and quantification of UCP1 protein in the BAT of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 4 per group). (K) Serum E level in male OXGR1OE AG mice treated with AKG for 12 weeks (n= 8 per group). (L-O). Oxygen consumption (L-M) and RER (N-O) of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 8 per group). (P-Q). Representative images (P) and quantification (Q) of gWAT and iWAT HE staining from male OXGR1OE AG mice treated with AKG for 12 weeks (n = 6 per group). (R-S). Representative images (R) and quantification (S) of p-HSL DAB staining from male OXGR1OE AG mice treated with AKG for 12 weeks (n = 6 per group). Results are presented as mean ± SEM. In (A), ** p≤0.01 by non-paired Student’s t test. In (B-C), *p≤0.05, **p≤0.01 by two-way ANOVA followed by post hoc Bonferroni tests. In (E-G), (I-K), (M), (O), (Q) and (S), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Turkey’s tests.

    Techniques Used: Over Expression, Expressing, Injection, Mouse Assay, Western Blot, Staining

    OXGR1 is required for metabolic beneficial effects of resistance exercise (A). Body weight gain in male WT littermates and OXGR1KO mice. At 8 weeks of age, male C57BL/6 WT control or OXGR1KO mice were switched to HFD. After 12 weeks of HFD feeding, mice were further divided into two groups, receiving non-exercise or resistance exercise for 14 days. (n = 8 per group). (B). Exercise-induced body weight loss in male WT littermates and OXGR1KO mice. Body weights from exercise mice were subtracted by the average body weight of non-exercise control group for each genotype (n = 8 per group). (C). Exercise-induced fat mass loss in male WT littermates and OXGR1KO mice. Fat mass from exercise mice were subtracted by the average fat mass of non-exercise control group for each genotype (n = 8 per group). (D). Cumulative food intake of male WT littermates and OXGR1KO mice after 14-day resistance exercise (n = 8 per group). (E-F). Weight index of gWAT (E) and iWAT (F) of male OXGR1KO mice after 14-days resistance exercise (n = 8 per group). (G). Body composition of male OXGR1KO mice after 14-days resistance exercise (n = 8 per group). (H). Serum AKG levels of male OXGR1KO mice after resistance exercise. Male OXGR1KO mice (10 weeks) fed with normal chow were receiving resistance exercise for 40 min (n = 8 per group). The serum AKG levels were tested before and immediately after exercise. (I). Serum E level in male OXGR1KO mice after 14-day resistance exercise (n = 8 per group). (J-K). The mRNA expression of UCP1 (J) in the BAT or HSL and ATGL (K) in the gWAT of male OXGR1KO mice after 14-day resistance exercise (n = 4 per group). (L-O). Oxygen consumption (L-M) and RER (N-O) in male OXGR1KO mice after 14-day resistance exercise (n = 8 per group). Results are presented as mean ± SEM. In (A-D) *p≤0.05, ** p≤0.01 by two-way ANOVA followed by post hoc Bonferroni tests. In (H), *p≤0.05 by non-paired Student’s t test. In (E-G), (I-K), (M) and (O), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Tukey’s tests.
    Figure Legend Snippet: OXGR1 is required for metabolic beneficial effects of resistance exercise (A). Body weight gain in male WT littermates and OXGR1KO mice. At 8 weeks of age, male C57BL/6 WT control or OXGR1KO mice were switched to HFD. After 12 weeks of HFD feeding, mice were further divided into two groups, receiving non-exercise or resistance exercise for 14 days. (n = 8 per group). (B). Exercise-induced body weight loss in male WT littermates and OXGR1KO mice. Body weights from exercise mice were subtracted by the average body weight of non-exercise control group for each genotype (n = 8 per group). (C). Exercise-induced fat mass loss in male WT littermates and OXGR1KO mice. Fat mass from exercise mice were subtracted by the average fat mass of non-exercise control group for each genotype (n = 8 per group). (D). Cumulative food intake of male WT littermates and OXGR1KO mice after 14-day resistance exercise (n = 8 per group). (E-F). Weight index of gWAT (E) and iWAT (F) of male OXGR1KO mice after 14-days resistance exercise (n = 8 per group). (G). Body composition of male OXGR1KO mice after 14-days resistance exercise (n = 8 per group). (H). Serum AKG levels of male OXGR1KO mice after resistance exercise. Male OXGR1KO mice (10 weeks) fed with normal chow were receiving resistance exercise for 40 min (n = 8 per group). The serum AKG levels were tested before and immediately after exercise. (I). Serum E level in male OXGR1KO mice after 14-day resistance exercise (n = 8 per group). (J-K). The mRNA expression of UCP1 (J) in the BAT or HSL and ATGL (K) in the gWAT of male OXGR1KO mice after 14-day resistance exercise (n = 4 per group). (L-O). Oxygen consumption (L-M) and RER (N-O) in male OXGR1KO mice after 14-day resistance exercise (n = 8 per group). Results are presented as mean ± SEM. In (A-D) *p≤0.05, ** p≤0.01 by two-way ANOVA followed by post hoc Bonferroni tests. In (H), *p≤0.05 by non-paired Student’s t test. In (E-G), (I-K), (M) and (O), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Tukey’s tests.

    Techniques Used: Mouse Assay, Expressing

    Acute in vivo effects of AKG (A-B). Serum levels of NE (A) and NEFA (B) in male C57BL/6 mice (10 weeks) 3 hrs after i.p. injection of saline or AKG (10 mg/kg) (n = 5-6 per group). (C). The mRNA expression of thermogenic genes in male C57BL/6 mice (10 weeks) 3 hrs after i.p. injection of saline or AKG (10 mg/kg) (n = 5-6 per group). (D). Immunoblots and quantification of UCP1 in BAT of male C57BL/6 mice (10 weeks) 3 hrs after i.p. injection of saline or AKG (10 mg/kg) (n = 3 per group). (E). Immunoblots and quantification of PLCβ and pErk in the adrenal glands of male C57BL/6 mice (10 weeks) 3 hrs after i.p. injection of saline or AKG (10 mg/kg) (n = 3 per group). (F-I). Physical activity (pedometer, F-G) and heart rate (H-I) of male mice i.p. injected with 10 mg/kg AKG or saline at 7:00 am (n = 8 per group). (J-L). Blood pressure of male mice i.p. inject ed with 10 mg/kg AKG or saline (n = 8 per group). Results are presented as mean ± SEM. In (A-E), (G), (I), (L), *p≤0.05, **p≤0.01, ***p≤0.01 by non-paired Student’s t-test. In (F), (H) and (J-K), *p≤0.05 by two-way ANOVA followed by post hoc Bonferroni tests.
    Figure Legend Snippet: Acute in vivo effects of AKG (A-B). Serum levels of NE (A) and NEFA (B) in male C57BL/6 mice (10 weeks) 3 hrs after i.p. injection of saline or AKG (10 mg/kg) (n = 5-6 per group). (C). The mRNA expression of thermogenic genes in male C57BL/6 mice (10 weeks) 3 hrs after i.p. injection of saline or AKG (10 mg/kg) (n = 5-6 per group). (D). Immunoblots and quantification of UCP1 in BAT of male C57BL/6 mice (10 weeks) 3 hrs after i.p. injection of saline or AKG (10 mg/kg) (n = 3 per group). (E). Immunoblots and quantification of PLCβ and pErk in the adrenal glands of male C57BL/6 mice (10 weeks) 3 hrs after i.p. injection of saline or AKG (10 mg/kg) (n = 3 per group). (F-I). Physical activity (pedometer, F-G) and heart rate (H-I) of male mice i.p. injected with 10 mg/kg AKG or saline at 7:00 am (n = 8 per group). (J-L). Blood pressure of male mice i.p. inject ed with 10 mg/kg AKG or saline (n = 8 per group). Results are presented as mean ± SEM. In (A-E), (G), (I), (L), *p≤0.05, **p≤0.01, ***p≤0.01 by non-paired Student’s t-test. In (F), (H) and (J-K), *p≤0.05 by two-way ANOVA followed by post hoc Bonferroni tests.

    Techniques Used: In Vivo, Mouse Assay, Injection, Expressing, Western Blot, Activity Assay

    AKG increases fat thermogenesis and lipolysis (A-D). Oxygen consumption (A-B) and respiratory exchange ratio (RER, C-D) in male C57BL/6 mice after 11 weeks of AKG supplementation (n = 8 per group). (E). Body temperature of male mice after 11 weeks of AKG supplementation (n = 9 per group). (F-G). Representative images (F) and quantification (G) of BAT thermogenesis induced by 6 hr cold exposure at 4℃ in male mice supplemented with AKG for 11 weeks (n = 9 per group). (H). The mRNA expression of PPARγ, FASN and ACC in the gWAT and iWAT from male mice supplemented with AKG for 11 weeks (n = 6 per group). (I-J). Immunoblots (I) and quantification (J) of p-HSL and ATGL protein in gWAT of male mice after 11 weeks of AKG supplementation (n = 3 per group). (K-L). DAB staining (K) and quantification (L) of p-HSL in gWAT and iWAT of male mice after 11 weeks of AKG supplementation (n = 9 per group). (M–O). The mRNA expression of thermogenic genes (M) and DAB staining (N) and quantification (O) of UCP1 in BAT of male mice supplemented with AKG for 11 weeks (n = 6-8 per group). (P). The mRNA expression of CD137, CD40, TBX1, TMEM26, CITED1 and slc27a1 in iWAT of male mice supplemented with AKG for 11 weeks (n = 8 per group). (Q–U). Serum levels of NEFA (Q), E (R), NE (S), T3 (T), and T4 (U) in male mice supplemented with AKG for 11 weeks (n = 8-9 per group). Results are presented as mean ± SEM. In (B), (D-E), (G-H), (J), (L-M), and (O-U), *p≤0.05, **p≤0.01, and ***p≤0.001 by non-paired Student’s t test.
    Figure Legend Snippet: AKG increases fat thermogenesis and lipolysis (A-D). Oxygen consumption (A-B) and respiratory exchange ratio (RER, C-D) in male C57BL/6 mice after 11 weeks of AKG supplementation (n = 8 per group). (E). Body temperature of male mice after 11 weeks of AKG supplementation (n = 9 per group). (F-G). Representative images (F) and quantification (G) of BAT thermogenesis induced by 6 hr cold exposure at 4℃ in male mice supplemented with AKG for 11 weeks (n = 9 per group). (H). The mRNA expression of PPARγ, FASN and ACC in the gWAT and iWAT from male mice supplemented with AKG for 11 weeks (n = 6 per group). (I-J). Immunoblots (I) and quantification (J) of p-HSL and ATGL protein in gWAT of male mice after 11 weeks of AKG supplementation (n = 3 per group). (K-L). DAB staining (K) and quantification (L) of p-HSL in gWAT and iWAT of male mice after 11 weeks of AKG supplementation (n = 9 per group). (M–O). The mRNA expression of thermogenic genes (M) and DAB staining (N) and quantification (O) of UCP1 in BAT of male mice supplemented with AKG for 11 weeks (n = 6-8 per group). (P). The mRNA expression of CD137, CD40, TBX1, TMEM26, CITED1 and slc27a1 in iWAT of male mice supplemented with AKG for 11 weeks (n = 8 per group). (Q–U). Serum levels of NEFA (Q), E (R), NE (S), T3 (T), and T4 (U) in male mice supplemented with AKG for 11 weeks (n = 8-9 per group). Results are presented as mean ± SEM. In (B), (D-E), (G-H), (J), (L-M), and (O-U), *p≤0.05, **p≤0.01, and ***p≤0.001 by non-paired Student’s t test.

    Techniques Used: Mouse Assay, Expressing, Western Blot, Staining

    Adrenal specific reexpression of OXGR1 rescues the stimulatory effects of AKG on thermogenesis and lipolysis (A). Serum E level in male OXGR1KO mice. At 12 weeks of age, male control or OXGR1KO mice were switched to HFD and received tap water or water supplemented with 2% AKG for 13 weeks (n = 8 per group). (B). Immunoblots and quantification of UCP1 protein expression in the BAT of male OXGR1KO mice treated with AKG for 13 weeks (n = 4 per group). (C-D). Representative images (C) and quantification (D) of iWAT and gWAT HE staining from male OXGR1KO mice treated with AKG for 13 weeks (n = 6 per group). (E-F). Representative images (E) and quantification (F) of p-HSL DAB staining from male OXGR1KO mice treated with AKG for 13 weeks (n = 6 per group). (G). The validation of OXGR1 reexpression. The mRNA expression of OXGR1 was determined in the adrenal glands from male WT control, OXGR1KO injected with HBAAV2/9-GFP, and OXGR1KO injected with HBAAV2/9-OXGR1 (OXGR1RE AG ) mice. (H). Serum E level in male OXGR1RE AG . Male OXGR1KO mice (8 weeks) were adrenal-specifically injected with control HBAAV2/9-GFP or HBAAV2/9-OXGR1. Two weeks after injections, mice were switched to HFD and further divided into two groups, receiving tap water or 2% AKG for 13 weeks. (n = 6 per group). (I). Immunoblots and quantification of UCP1 protein expression in the BAT of OXGR1RE AG mice treated with AKG for 13 weeks (n = 4 per group). (J-K). Representative images (J) and quantification (K) of iWAT and gWAT HE staining from OXGR1RE AG mice treated with AKG for 13 weeks (n = 6 per group). (L-M). Representative images (L) and quantification (M) of p-HSL DAB staining from OXGR1RE AG mice treated with AKG for 13 weeks (n = 6 per group). Results are presented as mean ± SEM. In (A-B), (D), (F), (H-I), (K) and (M), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Turkey’s tests.
    Figure Legend Snippet: Adrenal specific reexpression of OXGR1 rescues the stimulatory effects of AKG on thermogenesis and lipolysis (A). Serum E level in male OXGR1KO mice. At 12 weeks of age, male control or OXGR1KO mice were switched to HFD and received tap water or water supplemented with 2% AKG for 13 weeks (n = 8 per group). (B). Immunoblots and quantification of UCP1 protein expression in the BAT of male OXGR1KO mice treated with AKG for 13 weeks (n = 4 per group). (C-D). Representative images (C) and quantification (D) of iWAT and gWAT HE staining from male OXGR1KO mice treated with AKG for 13 weeks (n = 6 per group). (E-F). Representative images (E) and quantification (F) of p-HSL DAB staining from male OXGR1KO mice treated with AKG for 13 weeks (n = 6 per group). (G). The validation of OXGR1 reexpression. The mRNA expression of OXGR1 was determined in the adrenal glands from male WT control, OXGR1KO injected with HBAAV2/9-GFP, and OXGR1KO injected with HBAAV2/9-OXGR1 (OXGR1RE AG ) mice. (H). Serum E level in male OXGR1RE AG . Male OXGR1KO mice (8 weeks) were adrenal-specifically injected with control HBAAV2/9-GFP or HBAAV2/9-OXGR1. Two weeks after injections, mice were switched to HFD and further divided into two groups, receiving tap water or 2% AKG for 13 weeks. (n = 6 per group). (I). Immunoblots and quantification of UCP1 protein expression in the BAT of OXGR1RE AG mice treated with AKG for 13 weeks (n = 4 per group). (J-K). Representative images (J) and quantification (K) of iWAT and gWAT HE staining from OXGR1RE AG mice treated with AKG for 13 weeks (n = 6 per group). (L-M). Representative images (L) and quantification (M) of p-HSL DAB staining from OXGR1RE AG mice treated with AKG for 13 weeks (n = 6 per group). Results are presented as mean ± SEM. In (A-B), (D), (F), (H-I), (K) and (M), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Turkey’s tests.

    Techniques Used: Mouse Assay, Western Blot, Expressing, Staining, Injection

    Metabolic effects of AKG is mediated by adrenergic stimulation of thermogenesis and lipolysis (A). Serum AKG concentration-time profile obtained from male C57BL/6 mice (10 weeks) fed with normal chow before or after i.p AKG (10 mg/kg body weight). The serum AKG level were tested at 0, 1, 2, 4 and 6 hrs after injection (n = 8 per group). (B-C). Representative images (B) and quantification (C) of BAT thermogenesis after 6 hr cold exposure at 4℃. Male C57BL/6 mice (10 weeks) were i.p. injected with 10 mg/kg AKG or saline and immediately exposed to cold stress at 4℃ (n = 8 per group). (D). Immunoblots and quantification of p-HSL and ATGL in the gWAT of male C57BL/6 mice (10 weeks) 3 hrs after i.p. injection of 10 mg/kg AKG or saline (n = 3 per group). (E). Serum E level in AKG treated male mice 3 hrs after i.p. injection (n = 8 per group). (F-I). Oxygen consumption (F-G) and RER (H-I) in male C57BL/6 mice (10 weeks) i.p. injected with saline, 10 mg/kg AKG, 1 mg/kg SR59230A (ADRB3 inhibitor) or AKG + SR59230A (n = 8 per group). All injections were performed at 7:00 am of second day. Data was summarized in bar graph (G and I) by light or dark cycle of second day. (J-N). Body weight gain (J), cumulative food intake (K), body composition (L), fat weight (M) and serum NEFA (N) of shame or adrenalectomized male C57BL/6 mice. Male mice were adrenalectomized at 8 weeks of age. Two weeks after surgeries, male mice were switched to HFD and given free access to tap water or 2% AKG for 9 weeks (n = 8 per group). (O-P). Representative images (O) and quantification (P) of BAT thermogenesis after 6h cold exposure at 4℃ in shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 8 per group). (Q). The mRNA expression of themogenic genes in the BAT of shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 6 per group). (R-S) Immunoblots (R) and quantification (S) of p-HSL and ATGL protein in the gWAT of shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 4 per group). (T-U). Immunoblots (T) and quantification (U) of UCP1 protein in the BAT of shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 4 per group). Results are presented as mean ± SEM. In (A), *p≤0.05 by non-paired Student’s t test compared with before injection. In (C-E), *p≤0.05, **p≤ 0.01 by non-paired Student’s t test. In (J-K), *p≤0.05 by two-way ANOVA followed by post hoc Bonferroni tests. In (G), (I), (L-N), (P-Q), (S) and (U), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Tukey’s tests.
    Figure Legend Snippet: Metabolic effects of AKG is mediated by adrenergic stimulation of thermogenesis and lipolysis (A). Serum AKG concentration-time profile obtained from male C57BL/6 mice (10 weeks) fed with normal chow before or after i.p AKG (10 mg/kg body weight). The serum AKG level were tested at 0, 1, 2, 4 and 6 hrs after injection (n = 8 per group). (B-C). Representative images (B) and quantification (C) of BAT thermogenesis after 6 hr cold exposure at 4℃. Male C57BL/6 mice (10 weeks) were i.p. injected with 10 mg/kg AKG or saline and immediately exposed to cold stress at 4℃ (n = 8 per group). (D). Immunoblots and quantification of p-HSL and ATGL in the gWAT of male C57BL/6 mice (10 weeks) 3 hrs after i.p. injection of 10 mg/kg AKG or saline (n = 3 per group). (E). Serum E level in AKG treated male mice 3 hrs after i.p. injection (n = 8 per group). (F-I). Oxygen consumption (F-G) and RER (H-I) in male C57BL/6 mice (10 weeks) i.p. injected with saline, 10 mg/kg AKG, 1 mg/kg SR59230A (ADRB3 inhibitor) or AKG + SR59230A (n = 8 per group). All injections were performed at 7:00 am of second day. Data was summarized in bar graph (G and I) by light or dark cycle of second day. (J-N). Body weight gain (J), cumulative food intake (K), body composition (L), fat weight (M) and serum NEFA (N) of shame or adrenalectomized male C57BL/6 mice. Male mice were adrenalectomized at 8 weeks of age. Two weeks after surgeries, male mice were switched to HFD and given free access to tap water or 2% AKG for 9 weeks (n = 8 per group). (O-P). Representative images (O) and quantification (P) of BAT thermogenesis after 6h cold exposure at 4℃ in shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 8 per group). (Q). The mRNA expression of themogenic genes in the BAT of shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 6 per group). (R-S) Immunoblots (R) and quantification (S) of p-HSL and ATGL protein in the gWAT of shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 4 per group). (T-U). Immunoblots (T) and quantification (U) of UCP1 protein in the BAT of shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 4 per group). Results are presented as mean ± SEM. In (A), *p≤0.05 by non-paired Student’s t test compared with before injection. In (C-E), *p≤0.05, **p≤ 0.01 by non-paired Student’s t test. In (J-K), *p≤0.05 by two-way ANOVA followed by post hoc Bonferroni tests. In (G), (I), (L-N), (P-Q), (S) and (U), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Tukey’s tests.

    Techniques Used: Concentration Assay, Mouse Assay, Injection, Western Blot, Expressing

    Metabolic effects of AKG in mice fed on chow. (A-B). Body weight gain (A) and cumulative food intake (B) of male C57BL/6 mice. At 12 weeks of age, chow-fed male mice were divided into two groups, receiving tap water or water supplemented with 2% AKG for 6 weeks (n = 8 per group). (C-D). Body composition (C) and tissue weight (D) of male mice treated with AKG for 6 weeks (n = 7-8 per group). (E-F). Representative images (E) and quantification (F) of gWAT HE staining from male mice treated with AKG for 6 weeks (n = 8 per group). (G-H). Body weight gain (G) and cumulative food intake (H) of female C57BL/6 mice. At 12 weeks of age, chow-fed female mice were divided into two groups, receiving tap water or water supplemented with 2% AKG for 11 weeks (n = 8 per group). (I-J). Body composition (C) and tissue weight (D) of female mice treated with AKG for 11 weeks (n = 8 per group). (K-L). Representative images (K) and quantification (L) of gWAT HE staining from female mice treated with AKG for 11 weeks (n = 8 per group). (M). The mRNA expression of thermogenic genes in BAT of male C57BL/6 mice supplemented with AKG for 6 weeks (n = 6 per group). (N-P). Immunoblots and quantification of UCP1 (N) and representative images of DAB staining (O) and quantification (P) of UCP1 in BAT of male mice supplemented with AKG for 6 weeks (n = 3-6 per group). (Q-U). Serum levels of NEFA (Q), E (R), NE (S), T3 (T), and T4 (U) in male mice supplemented with AKG for 6 weeks (n = 6 per group). Results are presented as mean ± SEM. In (A-B) and (G-H), *p≤0.05 by two-way ANOVA followed by post hoc Bonferroni tests. In (C-D), (F), (I-J), (L-N) and (P-U), *p≤0.05, **p≤0.01, ***p≤0.01 by non-paired Student’s t-test.
    Figure Legend Snippet: Metabolic effects of AKG in mice fed on chow. (A-B). Body weight gain (A) and cumulative food intake (B) of male C57BL/6 mice. At 12 weeks of age, chow-fed male mice were divided into two groups, receiving tap water or water supplemented with 2% AKG for 6 weeks (n = 8 per group). (C-D). Body composition (C) and tissue weight (D) of male mice treated with AKG for 6 weeks (n = 7-8 per group). (E-F). Representative images (E) and quantification (F) of gWAT HE staining from male mice treated with AKG for 6 weeks (n = 8 per group). (G-H). Body weight gain (G) and cumulative food intake (H) of female C57BL/6 mice. At 12 weeks of age, chow-fed female mice were divided into two groups, receiving tap water or water supplemented with 2% AKG for 11 weeks (n = 8 per group). (I-J). Body composition (C) and tissue weight (D) of female mice treated with AKG for 11 weeks (n = 8 per group). (K-L). Representative images (K) and quantification (L) of gWAT HE staining from female mice treated with AKG for 11 weeks (n = 8 per group). (M). The mRNA expression of thermogenic genes in BAT of male C57BL/6 mice supplemented with AKG for 6 weeks (n = 6 per group). (N-P). Immunoblots and quantification of UCP1 (N) and representative images of DAB staining (O) and quantification (P) of UCP1 in BAT of male mice supplemented with AKG for 6 weeks (n = 3-6 per group). (Q-U). Serum levels of NEFA (Q), E (R), NE (S), T3 (T), and T4 (U) in male mice supplemented with AKG for 6 weeks (n = 6 per group). Results are presented as mean ± SEM. In (A-B) and (G-H), *p≤0.05 by two-way ANOVA followed by post hoc Bonferroni tests. In (C-D), (F), (I-J), (L-N) and (P-U), *p≤0.05, **p≤0.01, ***p≤0.01 by non-paired Student’s t-test.

    Techniques Used: Mouse Assay, Staining, Expressing, Western Blot

    2) Product Images from "Estrogen/ERα signaling axis participates in osteoblast maturation via upregulating chromosomal and mitochondrial complex gene expressions"

    Article Title: Estrogen/ERα signaling axis participates in osteoblast maturation via upregulating chromosomal and mitochondrial complex gene expressions

    Journal: Oncotarget

    doi: 10.18632/oncotarget.23453

    Effects of estradiol on translocation of estrogen receptor alpha (ERα) to mitochondria Human osteoblast-like U2OS cells were exposed to 10 nM of estradiol for 1, 6, 12, and 24 h. Distribution of the ERα protein in human osteoblasts was immunodetected using an antibody with Cy3-conjugated streptavidin ( A , top panel). Mitochondria of human osteoblasts were stained with 3,3′-dihexyloxacarbocyanine (DiOC6), a positively charged dye (middle panel). Merged signals indicated that the ERα protein had been translocated into mitochondria (bottom panels). These fluorescent signals were quantified and statistically analyzed (B) . Each value represents the mean ± SEM for n = 6. The symbol * indicates that the value significantly differed from the respective control group, p
    Figure Legend Snippet: Effects of estradiol on translocation of estrogen receptor alpha (ERα) to mitochondria Human osteoblast-like U2OS cells were exposed to 10 nM of estradiol for 1, 6, 12, and 24 h. Distribution of the ERα protein in human osteoblasts was immunodetected using an antibody with Cy3-conjugated streptavidin ( A , top panel). Mitochondria of human osteoblasts were stained with 3,3′-dihexyloxacarbocyanine (DiOC6), a positively charged dye (middle panel). Merged signals indicated that the ERα protein had been translocated into mitochondria (bottom panels). These fluorescent signals were quantified and statistically analyzed (B) . Each value represents the mean ± SEM for n = 6. The symbol * indicates that the value significantly differed from the respective control group, p

    Techniques Used: Translocation Assay, Staining

    Effects of estradiol on translocation of estrogen receptor alpha (ERα) to nuclei Human osteoblast-like U2OS cells were exposed to 10 nM of estradiol for 1, 6, 12, and 24 h. Distribution of the ERα protein in human osteoblasts was immunodetected using an antibody with Cy3-conjugated streptavidin ( A , top panel). Cellular nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI) (middle panel). The merged signals indicated that the ERα protein had been translocated into nuclei (bottom panel). These merged fluorescent signals were quantified and statistically analyzed (B) . Each value represents the mean ± SEM for n = 6. The symbol * indicates that the value significantly differed from the respective control group, p
    Figure Legend Snippet: Effects of estradiol on translocation of estrogen receptor alpha (ERα) to nuclei Human osteoblast-like U2OS cells were exposed to 10 nM of estradiol for 1, 6, 12, and 24 h. Distribution of the ERα protein in human osteoblasts was immunodetected using an antibody with Cy3-conjugated streptavidin ( A , top panel). Cellular nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI) (middle panel). The merged signals indicated that the ERα protein had been translocated into nuclei (bottom panel). These merged fluorescent signals were quantified and statistically analyzed (B) . Each value represents the mean ± SEM for n = 6. The symbol * indicates that the value significantly differed from the respective control group, p

    Techniques Used: Translocation Assay, Staining

    3) Product Images from "Exercise‐induced α‐ketoglutaric acid stimulates muscle hypertrophy and fat loss through OXGR1‐dependent adrenal activation"

    Article Title: Exercise‐induced α‐ketoglutaric acid stimulates muscle hypertrophy and fat loss through OXGR1‐dependent adrenal activation

    Journal: The EMBO Journal

    doi: 10.15252/embj.2019103304

    AKG increases fat thermogenesis and lipolysis Oxygen consumption in male C57BL/6 mice after 11 weeks of AKG supplementation ( n = 8 per group). Body temperature of male mice after 11 weeks of AKG supplementation ( n = 9 per group). Representative images (D) and quantification (E) of BAT thermogenesis induced by 6‐h cold exposure at 4°C in male mice supplemented with AKG for 11 weeks ( n = 9 per group). The mRNA expression of thermogenic genes (F) and immunoblots and quantification (G) of UCP1 protein in BAT of male mice after 11 weeks of AKG supplementation ( n = 3–6 per group). DAB staining (H) and quantification (I) of UCP1 in BAT of male mice supplemented with AKG for 11 weeks ( n = 9 per group). Serum levels of NEFA in male mice supplemented with AKG for 11 weeks ( n = 9 per group). The mRNA expression of CD137, CD40, TBX1, TMEM26, CITED1, and slc27a1 in iWAT of male mice supplemented with AKG for 11 weeks ( n = 8 per group). Respiratory exchange ratio (RER) in male C57BL/6 mice after 11 weeks of AKG supplementation ( n = 8 per group). Immunoblots (N) and quantification (O) of p‐HSL and ATGL protein in gWAT of male mice after 11 weeks of AKG supplementation ( n = 3 per group). Representative images (P) and quantification (Q) of p‐HSL DAB staining in gWAT and iWAT of male mice after 11 weeks of AKG supplementation ( n = 9 per group). The mRNA expression of PPARγ, FASN, and ACC in the gWAT and iWAT from male mice supplemented with AKG for 11 weeks ( n = 6 per group). Serum levels of E (S), NE (T), T3 (U), and T4 (V) in male mice supplemented with AKG for 11 weeks ( n = 8–9 per group). Data information: Results are presented as mean ± SEM. In (B, C, E–G, I–K, M, O and Q–V), * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001 by non‐paired Student's t ‐test.
    Figure Legend Snippet: AKG increases fat thermogenesis and lipolysis Oxygen consumption in male C57BL/6 mice after 11 weeks of AKG supplementation ( n = 8 per group). Body temperature of male mice after 11 weeks of AKG supplementation ( n = 9 per group). Representative images (D) and quantification (E) of BAT thermogenesis induced by 6‐h cold exposure at 4°C in male mice supplemented with AKG for 11 weeks ( n = 9 per group). The mRNA expression of thermogenic genes (F) and immunoblots and quantification (G) of UCP1 protein in BAT of male mice after 11 weeks of AKG supplementation ( n = 3–6 per group). DAB staining (H) and quantification (I) of UCP1 in BAT of male mice supplemented with AKG for 11 weeks ( n = 9 per group). Serum levels of NEFA in male mice supplemented with AKG for 11 weeks ( n = 9 per group). The mRNA expression of CD137, CD40, TBX1, TMEM26, CITED1, and slc27a1 in iWAT of male mice supplemented with AKG for 11 weeks ( n = 8 per group). Respiratory exchange ratio (RER) in male C57BL/6 mice after 11 weeks of AKG supplementation ( n = 8 per group). Immunoblots (N) and quantification (O) of p‐HSL and ATGL protein in gWAT of male mice after 11 weeks of AKG supplementation ( n = 3 per group). Representative images (P) and quantification (Q) of p‐HSL DAB staining in gWAT and iWAT of male mice after 11 weeks of AKG supplementation ( n = 9 per group). The mRNA expression of PPARγ, FASN, and ACC in the gWAT and iWAT from male mice supplemented with AKG for 11 weeks ( n = 6 per group). Serum levels of E (S), NE (T), T3 (U), and T4 (V) in male mice supplemented with AKG for 11 weeks ( n = 8–9 per group). Data information: Results are presented as mean ± SEM. In (B, C, E–G, I–K, M, O and Q–V), * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001 by non‐paired Student's t ‐test.

    Techniques Used: Mouse Assay, Expressing, Western Blot, Staining

    OXGR1 is required for metabolic beneficial effects of resistance exercise Body weight gain in male WT littermates and OXGR1KO mice. At 8 weeks of age, male C57BL/6 WT control or OXGR1KO mice were switched to HFD. After 12 weeks of HFD feeding, mice were further divided into two groups receiving non‐exercise or resistance exercise for 14 days ( n = 8 per group). Exercise‐induced body weight loss in male WT littermates and OXGR1KO mice. Body weights from exercise mice were subtracted by the average body weight of the non‐exercise control group for each genotype ( n = 8 per group). Exercise‐induced fat mass loss in male WT littermates and OXGR1KO mice. Fat mass from exercise mice was subtracted by the average fat mass of the non‐exercise control group for each genotype ( n = 8 per group). Cumulative food intake of male WT littermates and OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Weight index of gWAT (E) and iWAT (F) of male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Body composition of male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Serum AKG levels of male WT and OXGR1KO mice after resistance exercise. Male WT and OXGR1KO mice (10 weeks old) fed with normal chow were receiving resistance exercise for 40 min ( n = 8 per group). The serum AKG levels were tested before and immediately after exercise. Serum E level in male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). The mRNA expression (J) and protein expression of UCP1 (K) in the BAT or the mRNA expression of HSL and ATGL (L) in the gWAT of male OXGR1KO mice after 14‐day resistance exercise ( n = 4 per group). Oxygen consumption (M, N) and RER (O, P) in male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Data information: Results are presented as mean ± SEM. In (A–D) * P ≤ 0.05 and ** P ≤ 0.01 by two‐way ANOVA followed by post hoc Bonferroni tests. In (E–L, N and P), different letters between bars indicate P ≤ 0.05 by one‐way ANOVA followed by post hoc Tukey's tests.
    Figure Legend Snippet: OXGR1 is required for metabolic beneficial effects of resistance exercise Body weight gain in male WT littermates and OXGR1KO mice. At 8 weeks of age, male C57BL/6 WT control or OXGR1KO mice were switched to HFD. After 12 weeks of HFD feeding, mice were further divided into two groups receiving non‐exercise or resistance exercise for 14 days ( n = 8 per group). Exercise‐induced body weight loss in male WT littermates and OXGR1KO mice. Body weights from exercise mice were subtracted by the average body weight of the non‐exercise control group for each genotype ( n = 8 per group). Exercise‐induced fat mass loss in male WT littermates and OXGR1KO mice. Fat mass from exercise mice was subtracted by the average fat mass of the non‐exercise control group for each genotype ( n = 8 per group). Cumulative food intake of male WT littermates and OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Weight index of gWAT (E) and iWAT (F) of male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Body composition of male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Serum AKG levels of male WT and OXGR1KO mice after resistance exercise. Male WT and OXGR1KO mice (10 weeks old) fed with normal chow were receiving resistance exercise for 40 min ( n = 8 per group). The serum AKG levels were tested before and immediately after exercise. Serum E level in male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). The mRNA expression (J) and protein expression of UCP1 (K) in the BAT or the mRNA expression of HSL and ATGL (L) in the gWAT of male OXGR1KO mice after 14‐day resistance exercise ( n = 4 per group). Oxygen consumption (M, N) and RER (O, P) in male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Data information: Results are presented as mean ± SEM. In (A–D) * P ≤ 0.05 and ** P ≤ 0.01 by two‐way ANOVA followed by post hoc Bonferroni tests. In (E–L, N and P), different letters between bars indicate P ≤ 0.05 by one‐way ANOVA followed by post hoc Tukey's tests.

    Techniques Used: Mouse Assay, Expressing

    Metabolic effects of AKG are mediated by adrenergic stimulation of thermogenesis and lipolysis Serum AKG concentration–time profile obtained from male C57BL/6 mice (10 weeks old) fed with normal chow before or after i.p AKG (10 mg/kg body weight). The serum AKG level was tested at 0, 1, 2, 4, and 6 h after injection ( n = 8 per group). Representative images (B) and quantification (C) of BAT thermogenesis after 6‐h cold exposure at 4°C. Male C57BL/6 mice (10 weeks old) were i.p. injected with 10 mg/kg AKG or saline and immediately exposed to cold stress at 4°C ( n = 8 per group). Immunoblots and quantification of p‐HSL and ATGL in the gWAT of male C57BL/6 mice (10 weeks old) 3 h after i.p. injection of 10 mg/kg AKG or saline ( n = 3 per group). Serum E level in AKG‐treated male mice 3 h after i.p. injection ( n = 8 per group). Oxygen consumption (F‐G) and RER (H‐I) in male C57BL/6 mice (10 weeks old) i.p. injected with saline, 10 mg/kg AKG, 1 mg/kg SR59230A (ADRB3 inhibitor), or AKG + SR59230A ( n = 8 per group). All injections were performed at 7:00 am of the second day. Data are summarized in bar graph (G and I) by light or dark cycle of the second day. Body weight gain (J), cumulative food intake (K), body composition (L), fat weight (M), and serum NEFA (N) of sham or adrenalectomized male C57BL/6 mice. Male mice were adrenalectomized at 8 weeks of age. Two weeks after surgeries, male mice were switched to HFD and given free access to tap water or 2% AKG for 9 weeks ( n = 8 per group). Representative images (O) and quantification (P) of BAT thermogenesis after 6‐h cold exposure at 4°C in sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 8 per group). The mRNA expression of thermogenic genes in the BAT of sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 6 per group). Immunoblots (R) and quantification (S) of p‐HSL and ATGL protein in the gWAT of sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 4 per group). Immunoblots (T) and quantification (U) of UCP1 protein in the BAT of sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 4 per group). Data information: Results are presented as mean ± SEM. In (A), * P ≤ 0.05 by non‐paired Student's t ‐test compared with before injection. In (C–E), * P ≤ 0.05 and ** P ≤ 0.01 by non‐paired Student's t ‐test. In (J, K), * P ≤ 0.05 by two‐way ANOVA followed by post hoc Bonferroni tests. In (G, I, L–N, P, Q, S and U), different letters between bars indicate P ≤ 0.05 by one‐way ANOVA followed by post hoc Tukey's tests. Source data are available online for this figure.
    Figure Legend Snippet: Metabolic effects of AKG are mediated by adrenergic stimulation of thermogenesis and lipolysis Serum AKG concentration–time profile obtained from male C57BL/6 mice (10 weeks old) fed with normal chow before or after i.p AKG (10 mg/kg body weight). The serum AKG level was tested at 0, 1, 2, 4, and 6 h after injection ( n = 8 per group). Representative images (B) and quantification (C) of BAT thermogenesis after 6‐h cold exposure at 4°C. Male C57BL/6 mice (10 weeks old) were i.p. injected with 10 mg/kg AKG or saline and immediately exposed to cold stress at 4°C ( n = 8 per group). Immunoblots and quantification of p‐HSL and ATGL in the gWAT of male C57BL/6 mice (10 weeks old) 3 h after i.p. injection of 10 mg/kg AKG or saline ( n = 3 per group). Serum E level in AKG‐treated male mice 3 h after i.p. injection ( n = 8 per group). Oxygen consumption (F‐G) and RER (H‐I) in male C57BL/6 mice (10 weeks old) i.p. injected with saline, 10 mg/kg AKG, 1 mg/kg SR59230A (ADRB3 inhibitor), or AKG + SR59230A ( n = 8 per group). All injections were performed at 7:00 am of the second day. Data are summarized in bar graph (G and I) by light or dark cycle of the second day. Body weight gain (J), cumulative food intake (K), body composition (L), fat weight (M), and serum NEFA (N) of sham or adrenalectomized male C57BL/6 mice. Male mice were adrenalectomized at 8 weeks of age. Two weeks after surgeries, male mice were switched to HFD and given free access to tap water or 2% AKG for 9 weeks ( n = 8 per group). Representative images (O) and quantification (P) of BAT thermogenesis after 6‐h cold exposure at 4°C in sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 8 per group). The mRNA expression of thermogenic genes in the BAT of sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 6 per group). Immunoblots (R) and quantification (S) of p‐HSL and ATGL protein in the gWAT of sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 4 per group). Immunoblots (T) and quantification (U) of UCP1 protein in the BAT of sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 4 per group). Data information: Results are presented as mean ± SEM. In (A), * P ≤ 0.05 by non‐paired Student's t ‐test compared with before injection. In (C–E), * P ≤ 0.05 and ** P ≤ 0.01 by non‐paired Student's t ‐test. In (J, K), * P ≤ 0.05 by two‐way ANOVA followed by post hoc Bonferroni tests. In (G, I, L–N, P, Q, S and U), different letters between bars indicate P ≤ 0.05 by one‐way ANOVA followed by post hoc Tukey's tests. Source data are available online for this figure.

    Techniques Used: Concentration Assay, Mouse Assay, Injection, Western Blot, Expressing

    4) Product Images from "Exercise‐induced α‐ketoglutaric acid stimulates muscle hypertrophy and fat loss through OXGR1‐dependent adrenal activation"

    Article Title: Exercise‐induced α‐ketoglutaric acid stimulates muscle hypertrophy and fat loss through OXGR1‐dependent adrenal activation

    Journal: The EMBO Journal

    doi: 10.15252/embj.2019103304

    Acute in vivo effects of AKG Serum levels of NE (A) and NEFA (B) in male C57BL/6 mice (10 weeks old) 3 h after i.p. injection of saline or AKG (10 mg/kg) ( n = 5–6 per group). The mRNA expression of thermogenic genes in male C57BL/6 mice (10 weeks old) 3 h after i.p. injection of saline or AKG (10 mg/kg) ( n = 5–6 per group). Immunoblots and quantification of UCP1 in BAT of male C57BL/6 mice (10 weeks old) 3 h after i.p. injection of saline or AKG (10 mg/kg) ( n = 3 per group). Immunoblots and quantification of PLCβ and p‐Erk in the adrenal glands of male C57BL/6 mice (10 weeks old) 3 h after i.p. injection of saline or AKG (10 mg/kg) ( n = 3 per group). Physical activity (pedometer; F, G) and heart rate (H, I) of male mice i.p. injected with 10 mg/kg AKG or saline at 7:00 am ( n = 8 per group). Blood pressure of male mice i.p. injected with 10 mg/kg AKG or saline ( n = 8 per group). Serum levels of succinate (SUC) (M), fumaric acid (FUMA) (N), pyruvic acid (Pyr) (O), oxaloacetic acid (OAA) (P), α‐ketoleucine (α‐kehex) (Q), alpha‐ketoisovaleric acid (α‐keval) (R), and 2‐hydroxy‐3‐methylbutyric acid (2H3MA) (S) in male C57BL/6 mice (10 weeks old) 3 h after i.p. injection of saline or AKG (10 mg/kg) ( n = 8 per group). Data information: Results are presented as mean ± SEM. In (A–E, G, I, L, N and Q), * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.01 by non‐paired Student's t ‐test. In (F, H, J and K), * P ≤ 0.05 by two‐way ANOVA followed by post hoc Bonferroni tests.
    Figure Legend Snippet: Acute in vivo effects of AKG Serum levels of NE (A) and NEFA (B) in male C57BL/6 mice (10 weeks old) 3 h after i.p. injection of saline or AKG (10 mg/kg) ( n = 5–6 per group). The mRNA expression of thermogenic genes in male C57BL/6 mice (10 weeks old) 3 h after i.p. injection of saline or AKG (10 mg/kg) ( n = 5–6 per group). Immunoblots and quantification of UCP1 in BAT of male C57BL/6 mice (10 weeks old) 3 h after i.p. injection of saline or AKG (10 mg/kg) ( n = 3 per group). Immunoblots and quantification of PLCβ and p‐Erk in the adrenal glands of male C57BL/6 mice (10 weeks old) 3 h after i.p. injection of saline or AKG (10 mg/kg) ( n = 3 per group). Physical activity (pedometer; F, G) and heart rate (H, I) of male mice i.p. injected with 10 mg/kg AKG or saline at 7:00 am ( n = 8 per group). Blood pressure of male mice i.p. injected with 10 mg/kg AKG or saline ( n = 8 per group). Serum levels of succinate (SUC) (M), fumaric acid (FUMA) (N), pyruvic acid (Pyr) (O), oxaloacetic acid (OAA) (P), α‐ketoleucine (α‐kehex) (Q), alpha‐ketoisovaleric acid (α‐keval) (R), and 2‐hydroxy‐3‐methylbutyric acid (2H3MA) (S) in male C57BL/6 mice (10 weeks old) 3 h after i.p. injection of saline or AKG (10 mg/kg) ( n = 8 per group). Data information: Results are presented as mean ± SEM. In (A–E, G, I, L, N and Q), * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.01 by non‐paired Student's t ‐test. In (F, H, J and K), * P ≤ 0.05 by two‐way ANOVA followed by post hoc Bonferroni tests.

    Techniques Used: In Vivo, Mouse Assay, Injection, Expressing, Western Blot, Activity Assay

    AKG increases fat thermogenesis and lipolysis Oxygen consumption in male C57BL/6 mice after 11 weeks of AKG supplementation ( n = 8 per group). Body temperature of male mice after 11 weeks of AKG supplementation ( n = 9 per group). Representative images (D) and quantification (E) of BAT thermogenesis induced by 6‐h cold exposure at 4°C in male mice supplemented with AKG for 11 weeks ( n = 9 per group). The mRNA expression of thermogenic genes (F) and immunoblots and quantification (G) of UCP1 protein in BAT of male mice after 11 weeks of AKG supplementation ( n = 3–6 per group). DAB staining (H) and quantification (I) of UCP1 in BAT of male mice supplemented with AKG for 11 weeks ( n = 9 per group). Serum levels of NEFA in male mice supplemented with AKG for 11 weeks ( n = 9 per group). The mRNA expression of CD137, CD40, TBX1, TMEM26, CITED1, and slc27a1 in iWAT of male mice supplemented with AKG for 11 weeks ( n = 8 per group). Respiratory exchange ratio (RER) in male C57BL/6 mice after 11 weeks of AKG supplementation ( n = 8 per group). Immunoblots (N) and quantification (O) of p‐HSL and ATGL protein in gWAT of male mice after 11 weeks of AKG supplementation ( n = 3 per group). Representative images (P) and quantification (Q) of p‐HSL DAB staining in gWAT and iWAT of male mice after 11 weeks of AKG supplementation ( n = 9 per group). The mRNA expression of PPARγ, FASN, and ACC in the gWAT and iWAT from male mice supplemented with AKG for 11 weeks ( n = 6 per group). Serum levels of E (S), NE (T), T3 (U), and T4 (V) in male mice supplemented with AKG for 11 weeks ( n = 8–9 per group). Data information: Results are presented as mean ± SEM. In (B, C, E–G, I–K, M, O and Q–V), * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001 by non‐paired Student's t ‐test.
    Figure Legend Snippet: AKG increases fat thermogenesis and lipolysis Oxygen consumption in male C57BL/6 mice after 11 weeks of AKG supplementation ( n = 8 per group). Body temperature of male mice after 11 weeks of AKG supplementation ( n = 9 per group). Representative images (D) and quantification (E) of BAT thermogenesis induced by 6‐h cold exposure at 4°C in male mice supplemented with AKG for 11 weeks ( n = 9 per group). The mRNA expression of thermogenic genes (F) and immunoblots and quantification (G) of UCP1 protein in BAT of male mice after 11 weeks of AKG supplementation ( n = 3–6 per group). DAB staining (H) and quantification (I) of UCP1 in BAT of male mice supplemented with AKG for 11 weeks ( n = 9 per group). Serum levels of NEFA in male mice supplemented with AKG for 11 weeks ( n = 9 per group). The mRNA expression of CD137, CD40, TBX1, TMEM26, CITED1, and slc27a1 in iWAT of male mice supplemented with AKG for 11 weeks ( n = 8 per group). Respiratory exchange ratio (RER) in male C57BL/6 mice after 11 weeks of AKG supplementation ( n = 8 per group). Immunoblots (N) and quantification (O) of p‐HSL and ATGL protein in gWAT of male mice after 11 weeks of AKG supplementation ( n = 3 per group). Representative images (P) and quantification (Q) of p‐HSL DAB staining in gWAT and iWAT of male mice after 11 weeks of AKG supplementation ( n = 9 per group). The mRNA expression of PPARγ, FASN, and ACC in the gWAT and iWAT from male mice supplemented with AKG for 11 weeks ( n = 6 per group). Serum levels of E (S), NE (T), T3 (U), and T4 (V) in male mice supplemented with AKG for 11 weeks ( n = 8–9 per group). Data information: Results are presented as mean ± SEM. In (B, C, E–G, I–K, M, O and Q–V), * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001 by non‐paired Student's t ‐test.

    Techniques Used: Mouse Assay, Expressing, Western Blot, Staining

    OXGR1 is required for metabolic beneficial effects of resistance exercise Body weight gain in male WT littermates and OXGR1KO mice. At 8 weeks of age, male C57BL/6 WT control or OXGR1KO mice were switched to HFD. After 12 weeks of HFD feeding, mice were further divided into two groups receiving non‐exercise or resistance exercise for 14 days ( n = 8 per group). Exercise‐induced body weight loss in male WT littermates and OXGR1KO mice. Body weights from exercise mice were subtracted by the average body weight of the non‐exercise control group for each genotype ( n = 8 per group). Exercise‐induced fat mass loss in male WT littermates and OXGR1KO mice. Fat mass from exercise mice was subtracted by the average fat mass of the non‐exercise control group for each genotype ( n = 8 per group). Cumulative food intake of male WT littermates and OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Weight index of gWAT (E) and iWAT (F) of male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Body composition of male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Serum AKG levels of male WT and OXGR1KO mice after resistance exercise. Male WT and OXGR1KO mice (10 weeks old) fed with normal chow were receiving resistance exercise for 40 min ( n = 8 per group). The serum AKG levels were tested before and immediately after exercise. Serum E level in male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). The mRNA expression (J) and protein expression of UCP1 (K) in the BAT or the mRNA expression of HSL and ATGL (L) in the gWAT of male OXGR1KO mice after 14‐day resistance exercise ( n = 4 per group). Oxygen consumption (M, N) and RER (O, P) in male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Data information: Results are presented as mean ± SEM. In (A–D) * P ≤ 0.05 and ** P ≤ 0.01 by two‐way ANOVA followed by post hoc Bonferroni tests. In (E–L, N and P), different letters between bars indicate P ≤ 0.05 by one‐way ANOVA followed by post hoc Tukey's tests.
    Figure Legend Snippet: OXGR1 is required for metabolic beneficial effects of resistance exercise Body weight gain in male WT littermates and OXGR1KO mice. At 8 weeks of age, male C57BL/6 WT control or OXGR1KO mice were switched to HFD. After 12 weeks of HFD feeding, mice were further divided into two groups receiving non‐exercise or resistance exercise for 14 days ( n = 8 per group). Exercise‐induced body weight loss in male WT littermates and OXGR1KO mice. Body weights from exercise mice were subtracted by the average body weight of the non‐exercise control group for each genotype ( n = 8 per group). Exercise‐induced fat mass loss in male WT littermates and OXGR1KO mice. Fat mass from exercise mice was subtracted by the average fat mass of the non‐exercise control group for each genotype ( n = 8 per group). Cumulative food intake of male WT littermates and OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Weight index of gWAT (E) and iWAT (F) of male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Body composition of male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Serum AKG levels of male WT and OXGR1KO mice after resistance exercise. Male WT and OXGR1KO mice (10 weeks old) fed with normal chow were receiving resistance exercise for 40 min ( n = 8 per group). The serum AKG levels were tested before and immediately after exercise. Serum E level in male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). The mRNA expression (J) and protein expression of UCP1 (K) in the BAT or the mRNA expression of HSL and ATGL (L) in the gWAT of male OXGR1KO mice after 14‐day resistance exercise ( n = 4 per group). Oxygen consumption (M, N) and RER (O, P) in male OXGR1KO mice after 14‐day resistance exercise ( n = 8 per group). Data information: Results are presented as mean ± SEM. In (A–D) * P ≤ 0.05 and ** P ≤ 0.01 by two‐way ANOVA followed by post hoc Bonferroni tests. In (E–L, N and P), different letters between bars indicate P ≤ 0.05 by one‐way ANOVA followed by post hoc Tukey's tests.

    Techniques Used: Mouse Assay, Expressing

    Metabolic effects of AKG are mediated by adrenergic stimulation of thermogenesis and lipolysis Serum AKG concentration–time profile obtained from male C57BL/6 mice (10 weeks old) fed with normal chow before or after i.p AKG (10 mg/kg body weight). The serum AKG level was tested at 0, 1, 2, 4, and 6 h after injection ( n = 8 per group). Representative images (B) and quantification (C) of BAT thermogenesis after 6‐h cold exposure at 4°C. Male C57BL/6 mice (10 weeks old) were i.p. injected with 10 mg/kg AKG or saline and immediately exposed to cold stress at 4°C ( n = 8 per group). Immunoblots and quantification of p‐HSL and ATGL in the gWAT of male C57BL/6 mice (10 weeks old) 3 h after i.p. injection of 10 mg/kg AKG or saline ( n = 3 per group). Serum E level in AKG‐treated male mice 3 h after i.p. injection ( n = 8 per group). Oxygen consumption (F‐G) and RER (H‐I) in male C57BL/6 mice (10 weeks old) i.p. injected with saline, 10 mg/kg AKG, 1 mg/kg SR59230A (ADRB3 inhibitor), or AKG + SR59230A ( n = 8 per group). All injections were performed at 7:00 am of the second day. Data are summarized in bar graph (G and I) by light or dark cycle of the second day. Body weight gain (J), cumulative food intake (K), body composition (L), fat weight (M), and serum NEFA (N) of sham or adrenalectomized male C57BL/6 mice. Male mice were adrenalectomized at 8 weeks of age. Two weeks after surgeries, male mice were switched to HFD and given free access to tap water or 2% AKG for 9 weeks ( n = 8 per group). Representative images (O) and quantification (P) of BAT thermogenesis after 6‐h cold exposure at 4°C in sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 8 per group). The mRNA expression of thermogenic genes in the BAT of sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 6 per group). Immunoblots (R) and quantification (S) of p‐HSL and ATGL protein in the gWAT of sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 4 per group). Immunoblots (T) and quantification (U) of UCP1 protein in the BAT of sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 4 per group). Data information: Results are presented as mean ± SEM. In (A), * P ≤ 0.05 by non‐paired Student's t ‐test compared with before injection. In (C–E), * P ≤ 0.05 and ** P ≤ 0.01 by non‐paired Student's t ‐test. In (J, K), * P ≤ 0.05 by two‐way ANOVA followed by post hoc Bonferroni tests. In (G, I, L–N, P, Q, S and U), different letters between bars indicate P ≤ 0.05 by one‐way ANOVA followed by post hoc Tukey's tests. Source data are available online for this figure.
    Figure Legend Snippet: Metabolic effects of AKG are mediated by adrenergic stimulation of thermogenesis and lipolysis Serum AKG concentration–time profile obtained from male C57BL/6 mice (10 weeks old) fed with normal chow before or after i.p AKG (10 mg/kg body weight). The serum AKG level was tested at 0, 1, 2, 4, and 6 h after injection ( n = 8 per group). Representative images (B) and quantification (C) of BAT thermogenesis after 6‐h cold exposure at 4°C. Male C57BL/6 mice (10 weeks old) were i.p. injected with 10 mg/kg AKG or saline and immediately exposed to cold stress at 4°C ( n = 8 per group). Immunoblots and quantification of p‐HSL and ATGL in the gWAT of male C57BL/6 mice (10 weeks old) 3 h after i.p. injection of 10 mg/kg AKG or saline ( n = 3 per group). Serum E level in AKG‐treated male mice 3 h after i.p. injection ( n = 8 per group). Oxygen consumption (F‐G) and RER (H‐I) in male C57BL/6 mice (10 weeks old) i.p. injected with saline, 10 mg/kg AKG, 1 mg/kg SR59230A (ADRB3 inhibitor), or AKG + SR59230A ( n = 8 per group). All injections were performed at 7:00 am of the second day. Data are summarized in bar graph (G and I) by light or dark cycle of the second day. Body weight gain (J), cumulative food intake (K), body composition (L), fat weight (M), and serum NEFA (N) of sham or adrenalectomized male C57BL/6 mice. Male mice were adrenalectomized at 8 weeks of age. Two weeks after surgeries, male mice were switched to HFD and given free access to tap water or 2% AKG for 9 weeks ( n = 8 per group). Representative images (O) and quantification (P) of BAT thermogenesis after 6‐h cold exposure at 4°C in sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 8 per group). The mRNA expression of thermogenic genes in the BAT of sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 6 per group). Immunoblots (R) and quantification (S) of p‐HSL and ATGL protein in the gWAT of sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 4 per group). Immunoblots (T) and quantification (U) of UCP1 protein in the BAT of sham or adrenalectomized male mice treated with AKG for 9 weeks ( n = 4 per group). Data information: Results are presented as mean ± SEM. In (A), * P ≤ 0.05 by non‐paired Student's t ‐test compared with before injection. In (C–E), * P ≤ 0.05 and ** P ≤ 0.01 by non‐paired Student's t ‐test. In (J, K), * P ≤ 0.05 by two‐way ANOVA followed by post hoc Bonferroni tests. In (G, I, L–N, P, Q, S and U), different letters between bars indicate P ≤ 0.05 by one‐way ANOVA followed by post hoc Tukey's tests. Source data are available online for this figure.

    Techniques Used: Concentration Assay, Mouse Assay, Injection, Western Blot, Expressing

    Metabolic effects of AKG in mice fed on chow Body weight gain (A) and cumulative food intake (B) of male C57BL/6 mice. At 12 weeks of age, chow‐fed male mice were divided into two groups receiving tap water or water supplemented with 2% AKG for 6 weeks ( n = 8 per group). Body composition (C) and tissue weight (D) of male mice treated with AKG for 6 weeks ( n = 7–8 per group). Representative images (E) and quantification (F) of gWAT HE staining from male mice treated with AKG for 6 weeks ( n = 8 per group). Body weight gain (G) and cumulative food intake (H) of female C57BL/6 mice. At 12 weeks of age, chow‐fed female mice were divided into two groups receiving tap water or water supplemented with 2% AKG for 11 weeks ( n = 8 per group). Body composition (I) and tissue weight (J) of female mice treated with AKG for 11 weeks ( n = 8 per group). Representative images (K) and quantification (L) of gWAT HE staining from female mice treated with AKG for 11 weeks ( n = 8 per group). The mRNA expression of thermogenic genes in BAT of male C57BL/6 mice supplemented with AKG for 6 weeks ( n = 6 per group). Immunoblots and quantification of UCP1 (N) and representative images of DAB staining (O) and quantification (P) of UCP1 in BAT of male mice supplemented with AKG for 6 weeks ( n = 3–6 per group). Serum levels of NEFA (Q), E (R), NE (S), T3 (T), and T4 (U) in male mice supplemented with AKG for 6 weeks ( n = 6 per group). Data information: Results are presented as mean ± SEM. In (A, B, G and H), * P ≤ 0.05 by two‐way ANOVA followed by post hoc Bonferroni tests. In (C, D, F, I, J, L–N and P–U), * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.01 by non‐paired Student's t ‐test. Source data are available online for this figure.
    Figure Legend Snippet: Metabolic effects of AKG in mice fed on chow Body weight gain (A) and cumulative food intake (B) of male C57BL/6 mice. At 12 weeks of age, chow‐fed male mice were divided into two groups receiving tap water or water supplemented with 2% AKG for 6 weeks ( n = 8 per group). Body composition (C) and tissue weight (D) of male mice treated with AKG for 6 weeks ( n = 7–8 per group). Representative images (E) and quantification (F) of gWAT HE staining from male mice treated with AKG for 6 weeks ( n = 8 per group). Body weight gain (G) and cumulative food intake (H) of female C57BL/6 mice. At 12 weeks of age, chow‐fed female mice were divided into two groups receiving tap water or water supplemented with 2% AKG for 11 weeks ( n = 8 per group). Body composition (I) and tissue weight (J) of female mice treated with AKG for 11 weeks ( n = 8 per group). Representative images (K) and quantification (L) of gWAT HE staining from female mice treated with AKG for 11 weeks ( n = 8 per group). The mRNA expression of thermogenic genes in BAT of male C57BL/6 mice supplemented with AKG for 6 weeks ( n = 6 per group). Immunoblots and quantification of UCP1 (N) and representative images of DAB staining (O) and quantification (P) of UCP1 in BAT of male mice supplemented with AKG for 6 weeks ( n = 3–6 per group). Serum levels of NEFA (Q), E (R), NE (S), T3 (T), and T4 (U) in male mice supplemented with AKG for 6 weeks ( n = 6 per group). Data information: Results are presented as mean ± SEM. In (A, B, G and H), * P ≤ 0.05 by two‐way ANOVA followed by post hoc Bonferroni tests. In (C, D, F, I, J, L–N and P–U), * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.01 by non‐paired Student's t ‐test. Source data are available online for this figure.

    Techniques Used: Mouse Assay, Staining, Expressing, Western Blot

    Effects of AKG supplementation on fecal microbiota composition and iWAT browning in male mice Fecal energy of male C57BL/6 mice after 1, 4, and 11 weeks of AKG supplementation. At 12 weeks of age, male C57BL/6 mice were switched to HFD and received tap water or water supplemented with 2% AKG for 11 weeks ( n = 9 per group). The fecal microbial composition in the phylum (B) and genus (C) in male C57BL/6 mice receiving 2% AKG water supplementation for 1 weeks (AKG1) or 4 weeks (AKG2) ( n = 5 per group). Community structure test by ANOSIM and ADONIS of beta diversity in the genus between groups. Immunoblots (E) and quantification (F) of UCP1 protein in the iWAT of male C57BL/6 mice. At 10 weeks of age, male C57BL/6 mice were switched from chow to HFD and divided into four groups receiving tap water + room temperature (RT, 23°C), tap water + cold exposure (6°C), 2% AKG supplementation + 23°C, and 2% AKG supplementation + 6°C for one week ( n = 8 per group). Representative images (G) and quantification (H, I) of HE staining or UCP1 staining in iWAT of male C57BL/6 mice ( n = 8 per group). Data information: Results are presented as mean ± SEM. In (A), data are analyzed by two‐way ANOVA followed by post hoc Bonferroni tests. In (F, H and I), different letters between bars indicate P ≤ 0.05 by one‐way ANOVA followed by post hoc Tukey's tests.
    Figure Legend Snippet: Effects of AKG supplementation on fecal microbiota composition and iWAT browning in male mice Fecal energy of male C57BL/6 mice after 1, 4, and 11 weeks of AKG supplementation. At 12 weeks of age, male C57BL/6 mice were switched to HFD and received tap water or water supplemented with 2% AKG for 11 weeks ( n = 9 per group). The fecal microbial composition in the phylum (B) and genus (C) in male C57BL/6 mice receiving 2% AKG water supplementation for 1 weeks (AKG1) or 4 weeks (AKG2) ( n = 5 per group). Community structure test by ANOSIM and ADONIS of beta diversity in the genus between groups. Immunoblots (E) and quantification (F) of UCP1 protein in the iWAT of male C57BL/6 mice. At 10 weeks of age, male C57BL/6 mice were switched from chow to HFD and divided into four groups receiving tap water + room temperature (RT, 23°C), tap water + cold exposure (6°C), 2% AKG supplementation + 23°C, and 2% AKG supplementation + 6°C for one week ( n = 8 per group). Representative images (G) and quantification (H, I) of HE staining or UCP1 staining in iWAT of male C57BL/6 mice ( n = 8 per group). Data information: Results are presented as mean ± SEM. In (A), data are analyzed by two‐way ANOVA followed by post hoc Bonferroni tests. In (F, H and I), different letters between bars indicate P ≤ 0.05 by one‐way ANOVA followed by post hoc Tukey's tests.

    Techniques Used: Mouse Assay, Western Blot, Staining

    5) Product Images from "Multiplex giant magnetoresistive biosensor microarrays identify interferon-associated autoantibodies in systemic lupus erythematosus"

    Article Title: Multiplex giant magnetoresistive biosensor microarrays identify interferon-associated autoantibodies in systemic lupus erythematosus

    Journal: Scientific Reports

    doi: 10.1038/srep27623

    GMR biosensor autoantigen microarrays. ( a ) Optical images of a GMR biosensor chip and a cartridge with a reaction well (left). The sensor chip measures 10 × 12 mm and consists of an array of 8 × 10 sensors (total 80 sensors). Each sensor size is 100 × 100 μm (right). ( b ) A schematic of assaying antibody reactivity to autoantigens (not to scale). (1) Autoantigens were printed on the surface of the chip’s sensors. (2) The sample was added to the reaction well, allowing antibodies to bind to their corresponding antigens. (3) After washing, species-specific, biotinylated anti-IgG antibodies were used as a secondary reagent. (4) Streptavidin-coated MNPs bind to the biotinylated detection antibodies, and the respective sensor detects stray field from the bound MNPs.
    Figure Legend Snippet: GMR biosensor autoantigen microarrays. ( a ) Optical images of a GMR biosensor chip and a cartridge with a reaction well (left). The sensor chip measures 10 × 12 mm and consists of an array of 8 × 10 sensors (total 80 sensors). Each sensor size is 100 × 100 μm (right). ( b ) A schematic of assaying antibody reactivity to autoantigens (not to scale). (1) Autoantigens were printed on the surface of the chip’s sensors. (2) The sample was added to the reaction well, allowing antibodies to bind to their corresponding antigens. (3) After washing, species-specific, biotinylated anti-IgG antibodies were used as a secondary reagent. (4) Streptavidin-coated MNPs bind to the biotinylated detection antibodies, and the respective sensor detects stray field from the bound MNPs.

    Techniques Used: Chromatin Immunoprecipitation

    6) Product Images from "Bystander suppression of collagen-induced arthritis in mice fed ovalbumin"

    Article Title: Bystander suppression of collagen-induced arthritis in mice fed ovalbumin

    Journal: Arthritis Research & Therapy

    doi: 10.1186/ar1150

    Effects on anti-bovine collagen type II (BCII) antibody responses in mice fed ovalbumin (OVA). Mice were fed OVA or a standard diet for 7 days. One week after the last day on the OVA diet, the mice were immunized with BCII or BCII mixed with OVA, emulsified in Freund's complete adjuvant. Three weeks later the mice were immunized again, with the same antigen(s) emulsified in Freund's incomplete adjuvant. Numbers of IgG anti-BCII antibody-forming spleen cells (AFCs) (a) , and serum IgG (b) , IgG 1 (c) , and IgG 2a (d) anti-BCII antibody activity, 1 week after booster immunization. Each symbol represents one mouse and the bars represent the median values. Data were compared using the Mann–Whitney U test. * P
    Figure Legend Snippet: Effects on anti-bovine collagen type II (BCII) antibody responses in mice fed ovalbumin (OVA). Mice were fed OVA or a standard diet for 7 days. One week after the last day on the OVA diet, the mice were immunized with BCII or BCII mixed with OVA, emulsified in Freund's complete adjuvant. Three weeks later the mice were immunized again, with the same antigen(s) emulsified in Freund's incomplete adjuvant. Numbers of IgG anti-BCII antibody-forming spleen cells (AFCs) (a) , and serum IgG (b) , IgG 1 (c) , and IgG 2a (d) anti-BCII antibody activity, 1 week after booster immunization. Each symbol represents one mouse and the bars represent the median values. Data were compared using the Mann–Whitney U test. * P

    Techniques Used: Mouse Assay, Activity Assay, MANN-WHITNEY

    Effects on ovalbumin (OVA)-specific immune responses in mice fed OVA. Mice were fed OVA or a standard diet for 7 days. One week after the last day on the OVA diet, the mice were immunized with OVA or OVA mixed with bovine collagen type II (BCII) emulsified in Freund's complete adjuvant. Three weeks later the mice were immunized again, with the same antigen(s) emulsified in Freund's incomplete adjuvant. (a) IgG anti-OVA antibody activity in serum 1 week after the booster immunization. Each circular symbol represents one mouse and the bars represent the median values. Data were compared using the Mann–Whitney U test. In vitro proliferation (b) and secretion of IFN-γ (c) , IL-4 (d) , and IL-10 (e) by splenocytes after OVA restimulation 1 week after the booster immunization. The proliferation results are presented as proliferation indexes (mean counts per minute [cpm] in triplicate cultures stimulated with OVA/mean cpm in triplicate control cultures). Bars represent mean ± standard error of the mean ( n = 7–9). * P
    Figure Legend Snippet: Effects on ovalbumin (OVA)-specific immune responses in mice fed OVA. Mice were fed OVA or a standard diet for 7 days. One week after the last day on the OVA diet, the mice were immunized with OVA or OVA mixed with bovine collagen type II (BCII) emulsified in Freund's complete adjuvant. Three weeks later the mice were immunized again, with the same antigen(s) emulsified in Freund's incomplete adjuvant. (a) IgG anti-OVA antibody activity in serum 1 week after the booster immunization. Each circular symbol represents one mouse and the bars represent the median values. Data were compared using the Mann–Whitney U test. In vitro proliferation (b) and secretion of IFN-γ (c) , IL-4 (d) , and IL-10 (e) by splenocytes after OVA restimulation 1 week after the booster immunization. The proliferation results are presented as proliferation indexes (mean counts per minute [cpm] in triplicate cultures stimulated with OVA/mean cpm in triplicate control cultures). Bars represent mean ± standard error of the mean ( n = 7–9). * P

    Techniques Used: Mouse Assay, Activity Assay, MANN-WHITNEY, In Vitro

    7) Product Images from "UDP-Glucuronosyltransferase 1a Enzymes Are Present and Active in the Mouse Blastocyst"

    Article Title: UDP-Glucuronosyltransferase 1a Enzymes Are Present and Active in the Mouse Blastocyst

    Journal: Drug Metabolism and Disposition

    doi: 10.1124/dmd.114.059766

    Confocal immunofluorescence analysis of Ugt expression and localization in blastocysts. (A) Exemplary images of blastocysts stained with pan-specific antibodies against Ugt1a and Ugt2b, with DAPI staining to show the cell nuclei. Strong Ugt1a signal is
    Figure Legend Snippet: Confocal immunofluorescence analysis of Ugt expression and localization in blastocysts. (A) Exemplary images of blastocysts stained with pan-specific antibodies against Ugt1a and Ugt2b, with DAPI staining to show the cell nuclei. Strong Ugt1a signal is

    Techniques Used: Immunofluorescence, Expressing, Staining

    8) Product Images from "Barriers in contribution of human mesenchymal stem cells to murine muscle regeneration"

    Article Title: Barriers in contribution of human mesenchymal stem cells to murine muscle regeneration

    Journal: World Journal of Experimental Medicine

    doi: 10.5493/wjem.v5.i2.140

    Pax7 + cells in regenerating skeletal muscle implants. A, B: Examples of single (A and B) and pairs of (B) Pax7 + cells (arrows) attached or positioned in close proximity to myofibers in a fresh mouse implant at 14 d after implantation. Scale bar is 50
    Figure Legend Snippet: Pax7 + cells in regenerating skeletal muscle implants. A, B: Examples of single (A and B) and pairs of (B) Pax7 + cells (arrows) attached or positioned in close proximity to myofibers in a fresh mouse implant at 14 d after implantation. Scale bar is 50

    Techniques Used:

    9) Product Images from "Brief Communication: Magnetic Immuno-Detection of SARS-CoV-2 specific Antibodies"

    Article Title: Brief Communication: Magnetic Immuno-Detection of SARS-CoV-2 specific Antibodies

    Journal: bioRxiv

    doi: 10.1101/2020.06.02.131102

    Proof-of-concept MInD-based SARS-CoV-2 specific antibody detection. IFCs were coated with commercial 2 μg·ml −1 SARS-CoV-2 spike protein peptide and blocked with BSA. A corresponding antibody was diluted either in PBS (black curves) or spiked in human serum (red curves) and applied to IFCs. Biotinylated secondary antibodies were added, followed by application of streptavidin-functionalized magnetic particles. Assay time of this preliminary MInD setup was 42 minutes (without column preparation). Limit of detection (LOD) was determined by help of non-linear hill fit. Each data point represents mean ± SD (n = 2).
    Figure Legend Snippet: Proof-of-concept MInD-based SARS-CoV-2 specific antibody detection. IFCs were coated with commercial 2 μg·ml −1 SARS-CoV-2 spike protein peptide and blocked with BSA. A corresponding antibody was diluted either in PBS (black curves) or spiked in human serum (red curves) and applied to IFCs. Biotinylated secondary antibodies were added, followed by application of streptavidin-functionalized magnetic particles. Assay time of this preliminary MInD setup was 42 minutes (without column preparation). Limit of detection (LOD) was determined by help of non-linear hill fit. Each data point represents mean ± SD (n = 2).

    Techniques Used:

    Proof-of-concept MInD assay setup using IFC coated with SARS-CoV-2 antigen. Assay steps and assay time are indicated. IFCs were coated with commercial SARS-CoV-2 S-protein peptide and blocked with BSA. Corresponding antibody was diluted either in PBS or spiked in human serum and applied to IFCs. Biotinylated secondary antibodies were added, followed by application of streptavidin-functionalized MNP. Finally, IFCs were inserted into the portable magnetic read-out device. Measuring signal can be correlated to the amount of antibody in the sample and antibody titer can be determined. Assay time of this preliminary MInD setup was 42 min which is approximately four times faster than ELISA (161 min).
    Figure Legend Snippet: Proof-of-concept MInD assay setup using IFC coated with SARS-CoV-2 antigen. Assay steps and assay time are indicated. IFCs were coated with commercial SARS-CoV-2 S-protein peptide and blocked with BSA. Corresponding antibody was diluted either in PBS or spiked in human serum and applied to IFCs. Biotinylated secondary antibodies were added, followed by application of streptavidin-functionalized MNP. Finally, IFCs were inserted into the portable magnetic read-out device. Measuring signal can be correlated to the amount of antibody in the sample and antibody titer can be determined. Assay time of this preliminary MInD setup was 42 min which is approximately four times faster than ELISA (161 min).

    Techniques Used: Antigen Assay, Enzyme-linked Immunosorbent Assay

    ELISA-based detection of SARS-CoV-2 spike protein specific antibody in PBS-buffer (black curve) or spiked in human serum (red curve). Antibody was diluted to concentrations ranging from 1.22 ng·ml −1 up to 5000 ng·ml −1 in each matrix and applied onto 2 ng·ml −1 SARS-CoV-2 spike protein peptide coated and BSA blocked microtiter plates. After addition of biotinylated secondary antibody, streptavidin-AP was applied. Limit of detection (LOD) was calculated using non-linear Hill fit (R2=0.997 for PBS-buffer and 0.996 in serum). Assay time of ELISA was 161 minutes. Each data point represents mean ± SD (n = 4 for PBS-buffer and n = 3 for serum).
    Figure Legend Snippet: ELISA-based detection of SARS-CoV-2 spike protein specific antibody in PBS-buffer (black curve) or spiked in human serum (red curve). Antibody was diluted to concentrations ranging from 1.22 ng·ml −1 up to 5000 ng·ml −1 in each matrix and applied onto 2 ng·ml −1 SARS-CoV-2 spike protein peptide coated and BSA blocked microtiter plates. After addition of biotinylated secondary antibody, streptavidin-AP was applied. Limit of detection (LOD) was calculated using non-linear Hill fit (R2=0.997 for PBS-buffer and 0.996 in serum). Assay time of ELISA was 161 minutes. Each data point represents mean ± SD (n = 4 for PBS-buffer and n = 3 for serum).

    Techniques Used: Enzyme-linked Immunosorbent Assay, Serum Assay

    10) Product Images from "Neuromyelitis optica study model based on chronic infusion of autoantibodies in rat cerebrospinal fluid"

    Article Title: Neuromyelitis optica study model based on chronic infusion of autoantibodies in rat cerebrospinal fluid

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-016-0577-8

    Axonal damage and loss in the spinal cord and optic nerve of the NMO-rat. a Axon injury detected in the NMO-rat compared to the Control-rat (rats infused with IgG AQP4+ 2 and IgG Control 2, D7) using neurofilament immunostaining: reduced number of axons detected as NF-M-positive spots in the white matter (WM); fragmentation and reduced axon thickness in the gray matter (GM). b Classification (10–20 to 100–140 μm 2 , ImageJ) and quantification (mean by field) of NF-M-positive spots in the spinal cord of the NMO-rats ( n = 6) compared to the Control-rats ( n = 6): loss of axons with 60–140 μm 2 sections in the NMO-rats (in cart: evaluation of the total axon number, p = 0.03). c Co-detection of myelin alteration (MBP in green ) and axonal loss (neurofilament NF-M subtype in red ) in the spinal cord of the NMO-rat compared to the Control-rat. d Increased expression of the NF-H phosphorylated form, a marker of axon injury, detected by Western blot (pNF-H/NF-H ratio; p = 0.04). e Axon fragmentation and loss in the optic nerve of the NMO-rats compared to the Control-rat detected by NF-M immunostaining. Scale bars = 20 μm
    Figure Legend Snippet: Axonal damage and loss in the spinal cord and optic nerve of the NMO-rat. a Axon injury detected in the NMO-rat compared to the Control-rat (rats infused with IgG AQP4+ 2 and IgG Control 2, D7) using neurofilament immunostaining: reduced number of axons detected as NF-M-positive spots in the white matter (WM); fragmentation and reduced axon thickness in the gray matter (GM). b Classification (10–20 to 100–140 μm 2 , ImageJ) and quantification (mean by field) of NF-M-positive spots in the spinal cord of the NMO-rats ( n = 6) compared to the Control-rats ( n = 6): loss of axons with 60–140 μm 2 sections in the NMO-rats (in cart: evaluation of the total axon number, p = 0.03). c Co-detection of myelin alteration (MBP in green ) and axonal loss (neurofilament NF-M subtype in red ) in the spinal cord of the NMO-rat compared to the Control-rat. d Increased expression of the NF-H phosphorylated form, a marker of axon injury, detected by Western blot (pNF-H/NF-H ratio; p = 0.04). e Axon fragmentation and loss in the optic nerve of the NMO-rats compared to the Control-rat detected by NF-M immunostaining. Scale bars = 20 μm

    Techniques Used: Immunostaining, Expressing, Marker, Western Blot

    11) Product Images from "The Syk-binding Ubiquitin Ligase c-Cbl Mediates Signaling-dependent B Cell Receptor Ubiquitination and B Cell Receptor-mediated Antigen Processing and Presentation *"

    Article Title: The Syk-binding Ubiquitin Ligase c-Cbl Mediates Signaling-dependent B Cell Receptor Ubiquitination and B Cell Receptor-mediated Antigen Processing and Presentation *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M112.357640

    c-Cbl mediates Ag·BCR ubiquitination and processing. Upper panel , B cells were pulsed with anti-human IgM-btn for the indicated time (minutes) at 37 °C. The cells were lysed and ubiquitinated ligand-BCR complexes isolated by ubiquitin
    Figure Legend Snippet: c-Cbl mediates Ag·BCR ubiquitination and processing. Upper panel , B cells were pulsed with anti-human IgM-btn for the indicated time (minutes) at 37 °C. The cells were lysed and ubiquitinated ligand-BCR complexes isolated by ubiquitin

    Techniques Used: Isolation

    12) Product Images from "The scaffolding function of the RLTPR protein explains its essential role for CD28 co-stimulation in mouse and human T cells"

    Article Title: The scaffolding function of the RLTPR protein explains its essential role for CD28 co-stimulation in mouse and human T cells

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20160579

    Th differentiation and B cell responses in mice deprived of functional RLTPR molecules. (A) Sorted naive CD4 + T cells (2 × 10 5 ) from mice of the specified genotype were stimulated for 5 d with anti-CD3 and -CD28 under Th1, Th2, or Th17 differentiating conditions. After 5 d of culture, the absolute number of IFN-γ + (Th1 condition), IL-4 + (Th2 condition), and IL-17 + (Th17 condition) CD4 + T cells was determined. Each dot corresponds to a mouse and the mean (horizontal bar) is indicated. (B) WT and Rltpr bas/bas mice were immunized intraperitoneally at day 0 and 14 with the T cell–dependent antigen TNP-KLH. The concentration of TNP-specific immunoglobulins of the indicated isotypes (IgG2a, IgG2b, and IgG1) were assessed in individual mice before and 21 d after immunization. (C) WT and Rltpr bas/bas mice were immunized with the T cell–independent antigen TNP-LPS, and the concentration of TNP-specific IgM was assessed in individual mice before and 7 d after immunization. (D) Splenic B cells from mice of the specified genotype were stimulated with F(ab)’ 2 goat anti–mouse IgM antibody in the presence or absence of anti-CD40 antibody, or LPS. After 4 d of culture, B cell proliferation was evaluated. Mean and SEM are shown. Data are representative of two independent experiments. In A–C, each dot corresponds to a mouse and the mean (horizontal bar) is indicated. **, P ≤ 0.01; ****, P ≤ 0.001; ns, nonsignificant. In D, two animals were used per genotype.
    Figure Legend Snippet: Th differentiation and B cell responses in mice deprived of functional RLTPR molecules. (A) Sorted naive CD4 + T cells (2 × 10 5 ) from mice of the specified genotype were stimulated for 5 d with anti-CD3 and -CD28 under Th1, Th2, or Th17 differentiating conditions. After 5 d of culture, the absolute number of IFN-γ + (Th1 condition), IL-4 + (Th2 condition), and IL-17 + (Th17 condition) CD4 + T cells was determined. Each dot corresponds to a mouse and the mean (horizontal bar) is indicated. (B) WT and Rltpr bas/bas mice were immunized intraperitoneally at day 0 and 14 with the T cell–dependent antigen TNP-KLH. The concentration of TNP-specific immunoglobulins of the indicated isotypes (IgG2a, IgG2b, and IgG1) were assessed in individual mice before and 21 d after immunization. (C) WT and Rltpr bas/bas mice were immunized with the T cell–independent antigen TNP-LPS, and the concentration of TNP-specific IgM was assessed in individual mice before and 7 d after immunization. (D) Splenic B cells from mice of the specified genotype were stimulated with F(ab)’ 2 goat anti–mouse IgM antibody in the presence or absence of anti-CD40 antibody, or LPS. After 4 d of culture, B cell proliferation was evaluated. Mean and SEM are shown. Data are representative of two independent experiments. In A–C, each dot corresponds to a mouse and the mean (horizontal bar) is indicated. **, P ≤ 0.01; ****, P ≤ 0.001; ns, nonsignificant. In D, two animals were used per genotype.

    Techniques Used: Mouse Assay, Functional Assay, Concentration Assay

    13) Product Images from "Gammaherpesvirus Latency Accentuates EAE Pathogenesis: Relevance to Epstein-Barr Virus and Multiple Sclerosis"

    Article Title: Gammaherpesvirus Latency Accentuates EAE Pathogenesis: Relevance to Epstein-Barr Virus and Multiple Sclerosis

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1002715

    γHV-68 EAE mice show increased levels of pro-inflammatory cytokines and chemokines and a decreased anti-MOG IgG1/IgG2a ratio in the serum. Mice were infected i.p. with γHV-68 (black bars) or MEM only (open bars). Five weeks p.i., EAE was induced. At day 10, 15 and 28 post EAE induction blood was harvested through a cardiac puncture and the levels of different ( A ) cytokines (day 28 shown here) and ( B ) chemokines (day 10 shown here) were evaluated using BD Cytometric Bead Array kits. Data from day 28 are shown for both IFN-γ and TNF-α; similar results were obtained at day 10 and 15 post EAE induction. Three-two separate experiments for each time point with 3–6 mice/group. Data were analyzed with t-test: *** p
    Figure Legend Snippet: γHV-68 EAE mice show increased levels of pro-inflammatory cytokines and chemokines and a decreased anti-MOG IgG1/IgG2a ratio in the serum. Mice were infected i.p. with γHV-68 (black bars) or MEM only (open bars). Five weeks p.i., EAE was induced. At day 10, 15 and 28 post EAE induction blood was harvested through a cardiac puncture and the levels of different ( A ) cytokines (day 28 shown here) and ( B ) chemokines (day 10 shown here) were evaluated using BD Cytometric Bead Array kits. Data from day 28 are shown for both IFN-γ and TNF-α; similar results were obtained at day 10 and 15 post EAE induction. Three-two separate experiments for each time point with 3–6 mice/group. Data were analyzed with t-test: *** p

    Techniques Used: Mouse Assay, Infection

    14) Product Images from "Participation of GATA-3 in regulation of bone healing through transcriptional upregulation of bcl-xL expression"

    Article Title: Participation of GATA-3 in regulation of bone healing through transcriptional upregulation of bcl-xL expression

    Journal: Experimental & Molecular Medicine

    doi: 10.1038/emm.2017.182

    Association of nuclear GATA-3 and Runt-related transcription factor 2 (Runx2) during bone healing. Male ICR mice were anesthetized, and a metaphyseal bone defect was created in the left proximal femurs. The right femurs were treated as the sham-operated group. At 1, 3, 5, and 7 days after surgery, the femurs were collected and cleaned. The expression of GATA-3 (green signals) and Runx2 (red signals) in the bone defect sites were analyzed using confocal microscopy ( a ). Nuclei were stained with DAPI (blue signals). The association of nuclear GATA-3 and Runx2 (white signals) was observed, quantified and statistically analyzed ( b ). The complex of GATA-3 associated with Runx2 was immunoprecipitated (IP) using an antibody against GATA-3, electrophoretically separated, and finally immunodetected using a Runx2 antibody ( c , top panel). Levels of GATA-3 and phosphorylated (p)GATA-3 were immunodetected (middle panel). The amount of IgG heavy chains (IgG-H) was determined as the internal standard (bottom panel). The association of GATA-3 and Runx2 in MC3T3-E1 osteoblasts (Ob) was assayed as the positive control. These protein bands were quantified and statistically analyzed ( d ). Each value represents the mean±s.e.m. for n =4. * Values significantly differ from the respective control group, P
    Figure Legend Snippet: Association of nuclear GATA-3 and Runt-related transcription factor 2 (Runx2) during bone healing. Male ICR mice were anesthetized, and a metaphyseal bone defect was created in the left proximal femurs. The right femurs were treated as the sham-operated group. At 1, 3, 5, and 7 days after surgery, the femurs were collected and cleaned. The expression of GATA-3 (green signals) and Runx2 (red signals) in the bone defect sites were analyzed using confocal microscopy ( a ). Nuclei were stained with DAPI (blue signals). The association of nuclear GATA-3 and Runx2 (white signals) was observed, quantified and statistically analyzed ( b ). The complex of GATA-3 associated with Runx2 was immunoprecipitated (IP) using an antibody against GATA-3, electrophoretically separated, and finally immunodetected using a Runx2 antibody ( c , top panel). Levels of GATA-3 and phosphorylated (p)GATA-3 were immunodetected (middle panel). The amount of IgG heavy chains (IgG-H) was determined as the internal standard (bottom panel). The association of GATA-3 and Runx2 in MC3T3-E1 osteoblasts (Ob) was assayed as the positive control. These protein bands were quantified and statistically analyzed ( d ). Each value represents the mean±s.e.m. for n =4. * Values significantly differ from the respective control group, P

    Techniques Used: Mouse Assay, Expressing, Confocal Microscopy, Staining, Immunoprecipitation, Positive Control

    15) Product Images from "Combinatorial synthesis and screening of cancer cell-specific nanomedicines targeted via phage fusion proteins"

    Article Title: Combinatorial synthesis and screening of cancer cell-specific nanomedicines targeted via phage fusion proteins

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2015.00628

    Purification and characterization of DMPGTVLP-modified Lipodox . Size exclusion chromatography of modified Lipodox on a Superose 6 column (30 × 1 cm), 2.5 mL/fractions 4–11 of the elution profile are shown. Western blot of recovered fractions probed with a polyclonal anti-fd rabbit primary IgG, followed by a biotinylated goat anti-rabbit secondary IgG and detected with NeutrAvadin-HRP and Pico West luminol substrate. Physicochemical characterization of recovered fractions for size distribution (d.nm), zeta potential (mV), doxorubicin recovery (μg), and percent doxorubicin recovery (%).
    Figure Legend Snippet: Purification and characterization of DMPGTVLP-modified Lipodox . Size exclusion chromatography of modified Lipodox on a Superose 6 column (30 × 1 cm), 2.5 mL/fractions 4–11 of the elution profile are shown. Western blot of recovered fractions probed with a polyclonal anti-fd rabbit primary IgG, followed by a biotinylated goat anti-rabbit secondary IgG and detected with NeutrAvadin-HRP and Pico West luminol substrate. Physicochemical characterization of recovered fractions for size distribution (d.nm), zeta potential (mV), doxorubicin recovery (μg), and percent doxorubicin recovery (%).

    Techniques Used: Purification, Modification, Size-exclusion Chromatography, Western Blot

    16) Product Images from "Characterization of novel biomarkers in selecting for subtype specific medulloblastoma phenotypes"

    Article Title: Characterization of novel biomarkers in selecting for subtype specific medulloblastoma phenotypes

    Journal: Oncotarget

    doi:

    CD271 levels are inversely correlated with expression of SHH pathway genes A. - C. qPCR analysis of PTCH , SMO , GLI1 and GLI2 gene expression in Daoy sorted CD271+ vs . CD271− cells (A), MED311 sorted CD271+ vs . CD271− cells (B), and Daoy OE vs . negative control cells (C). ( D. - E.) qPCR analysis of PTCH , SMO , GLI1 and GLI2 gene expression in UI226 (D) and MED311 (E) cells following knockdown of CD271. Error bars: s.e.m. P
    Figure Legend Snippet: CD271 levels are inversely correlated with expression of SHH pathway genes A. - C. qPCR analysis of PTCH , SMO , GLI1 and GLI2 gene expression in Daoy sorted CD271+ vs . CD271− cells (A), MED311 sorted CD271+ vs . CD271− cells (B), and Daoy OE vs . negative control cells (C). ( D. - E.) qPCR analysis of PTCH , SMO , GLI1 and GLI2 gene expression in UI226 (D) and MED311 (E) cells following knockdown of CD271. Error bars: s.e.m. P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Negative Control

    Candidate cell surface markers are differentially expressed in MB cell lines/primary cultures and patient samples A. Representative dot plots of staining for candidate biomarkers in Daoy, MED311-FH and UI226 tumorspheres (SHH variant) vs . tumorspheres from Group 3/Group 4 cell lines by flow cytometry. Insets: respective isotype controls. 7AAD: 7-aminoactinomycin D cell viability dye. B. - D. CD271 (B), CD171 (C) and CD106 (D) expression in paraffin embedded sections of fetal cerebellum and primary medulloblastoma samples. Scale bar: 400 μm.
    Figure Legend Snippet: Candidate cell surface markers are differentially expressed in MB cell lines/primary cultures and patient samples A. Representative dot plots of staining for candidate biomarkers in Daoy, MED311-FH and UI226 tumorspheres (SHH variant) vs . tumorspheres from Group 3/Group 4 cell lines by flow cytometry. Insets: respective isotype controls. 7AAD: 7-aminoactinomycin D cell viability dye. B. - D. CD271 (B), CD171 (C) and CD106 (D) expression in paraffin embedded sections of fetal cerebellum and primary medulloblastoma samples. Scale bar: 400 μm.

    Techniques Used: Staining, Variant Assay, Flow Cytometry, Cytometry, Expressing

    CD271 overexpression changes the size and number of Daoy tumorspheres A. pReceiver-Lv105 lentiviral construct used for stable overexpression of CD271 in Daoy cells. B. - C. Validation of CD271 OE in Daoy cells by flow cytometry (B) and Western blot (C). GAPDH serves as a loading control. D. - F. Representative images of primary (D), secondary (E), and tertiary (F) tumorspheres from Daoy negative control cells and stable Daoy CD271 OE cells. G. - H. Primary tumorsphere number (G) and tumorsphere size (H) are increased in Daoy OEs vs . controls I. - L. Tumorsphere number is decreased in secondary (I) and tertiary (K) Daoy OE cells compared to controls; whereas tumorsphere size is increased in secondary (J) and tertiary Daoy CD271 OEs (L). Error bars: s.e.m. P
    Figure Legend Snippet: CD271 overexpression changes the size and number of Daoy tumorspheres A. pReceiver-Lv105 lentiviral construct used for stable overexpression of CD271 in Daoy cells. B. - C. Validation of CD271 OE in Daoy cells by flow cytometry (B) and Western blot (C). GAPDH serves as a loading control. D. - F. Representative images of primary (D), secondary (E), and tertiary (F) tumorspheres from Daoy negative control cells and stable Daoy CD271 OE cells. G. - H. Primary tumorsphere number (G) and tumorsphere size (H) are increased in Daoy OEs vs . controls I. - L. Tumorsphere number is decreased in secondary (I) and tertiary (K) Daoy OE cells compared to controls; whereas tumorsphere size is increased in secondary (J) and tertiary Daoy CD271 OEs (L). Error bars: s.e.m. P

    Techniques Used: Over Expression, Construct, Flow Cytometry, Cytometry, Western Blot, Negative Control

    γ- secretase inhibitor (Compound X) treatment of Daoy CD271 OE cells results in a reversal of the OE phenotype A. Western blot demonstrating C-terminal fragment accumulation upon blocking of CD271 cleavage by the γ- secretase inhibitor, Compound X. B. - E. Representative images of primary Daoy NEG (B-C) and Daoy CD271 OE (D-E) tumorspheres following treatment with Compound X. Scale bar: 400 μm. F. - G. Treatment of Daoy NEG and Daoy CD271 OE primary tumorspheres with Compound X results in an increase in sphere number (F) and decrease in sphere size (G). H. - I. Quantification of cell viability (H) and total cell number (I) in Daoy NEG and Daoy CD271 OE primary tumorspheres following treatment with Compound X. J. - M. Representative images of secondary Daoy NEG (J-K) and Daoy CD271 OE (L-M) tumorspheres following treatment with Compound X. Scale bar: 400 μm. N. - O. Treatment of Daoy NEG and Daoy CD271 OE secondary tumorspheres with Compound X either in passage 1 and 2 or in passage 1 only (Prev Tx). P. - Q. Quantification of cell viability (P) and total cell number (Q) in Daoy NEG and Daoy CD271 OE secondary tumorspheres following treatment with Compound X. Error bars: s.e.m. P
    Figure Legend Snippet: γ- secretase inhibitor (Compound X) treatment of Daoy CD271 OE cells results in a reversal of the OE phenotype A. Western blot demonstrating C-terminal fragment accumulation upon blocking of CD271 cleavage by the γ- secretase inhibitor, Compound X. B. - E. Representative images of primary Daoy NEG (B-C) and Daoy CD271 OE (D-E) tumorspheres following treatment with Compound X. Scale bar: 400 μm. F. - G. Treatment of Daoy NEG and Daoy CD271 OE primary tumorspheres with Compound X results in an increase in sphere number (F) and decrease in sphere size (G). H. - I. Quantification of cell viability (H) and total cell number (I) in Daoy NEG and Daoy CD271 OE primary tumorspheres following treatment with Compound X. J. - M. Representative images of secondary Daoy NEG (J-K) and Daoy CD271 OE (L-M) tumorspheres following treatment with Compound X. Scale bar: 400 μm. N. - O. Treatment of Daoy NEG and Daoy CD271 OE secondary tumorspheres with Compound X either in passage 1 and 2 or in passage 1 only (Prev Tx). P. - Q. Quantification of cell viability (P) and total cell number (Q) in Daoy NEG and Daoy CD271 OE secondary tumorspheres following treatment with Compound X. Error bars: s.e.m. P

    Techniques Used: Western Blot, Blocking Assay

    CD271 knockdown results in significantly smaller tumorspheres A. Validation of shRNA CD271 KD in Daoy OE cells by Western blot. GAPDH serves as a loading control. B. - C. Representative images of primary (B) and secondary (C) tumorspheres from Daoy OE cells infected with scrambled negative control vs . stable CD271 KD from 2 independent shRNA sequences. D. - E. Quantification of primary (D) and secondary (E) tumorsphere number and size following CD271 KD in Daoy CD271 OE cells. F. Validation of shRNA CD271 KD in UI226 cells by Western blot. GAPDH serves as a loading control. G. - H. Representative images of primary (G) and secondary (H) tumorspheres from UI226 cells infected with scrambled negative control vs . stable CD271 KD from 2 independent shRNA sequences. I. - J. Quantification of primary (I) and secondary (J) tumorsphere number and size following CD271 KD in UI226 cells. K. Validation of shRNA CD271 KD in MED311 cells by Western blot. GAPDH serves as a loading control. L. - M. Representative images of primary (L) and secondary (M) tumorspheres from MED311 cells infected with scrambled negative control vs . stable CD271 KD from 2 independent shRNA sequences. N. - O. Quantification of primary (N) and secondary (O) tumorsphere number and size following CD271 KD in MED311 cells. Error bars: s.e.m. P
    Figure Legend Snippet: CD271 knockdown results in significantly smaller tumorspheres A. Validation of shRNA CD271 KD in Daoy OE cells by Western blot. GAPDH serves as a loading control. B. - C. Representative images of primary (B) and secondary (C) tumorspheres from Daoy OE cells infected with scrambled negative control vs . stable CD271 KD from 2 independent shRNA sequences. D. - E. Quantification of primary (D) and secondary (E) tumorsphere number and size following CD271 KD in Daoy CD271 OE cells. F. Validation of shRNA CD271 KD in UI226 cells by Western blot. GAPDH serves as a loading control. G. - H. Representative images of primary (G) and secondary (H) tumorspheres from UI226 cells infected with scrambled negative control vs . stable CD271 KD from 2 independent shRNA sequences. I. - J. Quantification of primary (I) and secondary (J) tumorsphere number and size following CD271 KD in UI226 cells. K. Validation of shRNA CD271 KD in MED311 cells by Western blot. GAPDH serves as a loading control. L. - M. Representative images of primary (L) and secondary (M) tumorspheres from MED311 cells infected with scrambled negative control vs . stable CD271 KD from 2 independent shRNA sequences. N. - O. Quantification of primary (N) and secondary (O) tumorsphere number and size following CD271 KD in MED311 cells. Error bars: s.e.m. P

    Techniques Used: shRNA, Western Blot, Infection, Negative Control

    Transcript levels of CD271/p75NTR, CD171L1CAM, EGFR and CD106/VCAM1 across the 4 MB molecular variants Note that the CD271 Toronto dataset was previously published in Neoplasia, 15, Morrison et al., Deconstruction of Medulloblastoma cellular heterogeneity reveals differences between the most highly invasion and self-renewing phenotypes , 384-398, Copyright Elsevier (2013). A. - C. Gene expression data from 3 independent transcriptome datasets representing 548 patient samples showing downregulation of CD171/NCAM-L1 and relative enrichment of CD271/p75NTR, CD106/VCAM1, and EGFR, in SHH tumors compared with the other variants. Bars denote 1.5 interquartile range within each group. All subgroups were compared using a Kruskal-Wallis test for significance. Data are presented as log2 -transformed signal intensity. P
    Figure Legend Snippet: Transcript levels of CD271/p75NTR, CD171L1CAM, EGFR and CD106/VCAM1 across the 4 MB molecular variants Note that the CD271 Toronto dataset was previously published in Neoplasia, 15, Morrison et al., Deconstruction of Medulloblastoma cellular heterogeneity reveals differences between the most highly invasion and self-renewing phenotypes , 384-398, Copyright Elsevier (2013). A. - C. Gene expression data from 3 independent transcriptome datasets representing 548 patient samples showing downregulation of CD171/NCAM-L1 and relative enrichment of CD271/p75NTR, CD106/VCAM1, and EGFR, in SHH tumors compared with the other variants. Bars denote 1.5 interquartile range within each group. All subgroups were compared using a Kruskal-Wallis test for significance. Data are presented as log2 -transformed signal intensity. P

    Techniques Used: Expressing, Transformation Assay

    17) Product Images from "α-ketoglutaric acid stimulates muscle hypertrophy and fat loss through OXGR1-dependent adrenal activation"

    Article Title: α-ketoglutaric acid stimulates muscle hypertrophy and fat loss through OXGR1-dependent adrenal activation

    Journal: bioRxiv

    doi: 10.1101/796037

    Adrenal specific overexpression of OXGR1 enhances stimulatory effects of AKG on thermogenesis and lipolysis (A). The validation of OXGR1 overexpression. The mRNA expression of OXGR1 was determined in the adrenal glands from male WT control, WT injected with HBAAV2/9-GFP, and WT injected with HBAAV2/9-OXGR1 (OXGR1OE AG ) mice (n=5 per group). (B-C). Body weight gain (B) and cumulative food intake (C) of OXGR1OE AG . Male C57BL/6 mice (8 weeks) were adrenal-specifically injected with control HBAAV2/9-GFP or HBAAV2/9-OXGR1. Two weeks after injections, mice were switched to HFD and further divided into two groups, receiving tap water or water supplemented with 2% AKG for 12 weeks (n = 8 per group). (D-E). Representative image of body composition (D) and fat and lean mass index (E) of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 8 per group). (F-G). Weight index of gWAT (F) and iWAT (G) in male OXGR1OE AG mice treated with AKG for 12 weeks (n = 6 per group). (H-I). Immunoblots (H) and quantification (I) of p-HSL and ATGL protein in the gWAT of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 4 per group). (J). Immunoblots and quantification of UCP1 protein in the BAT of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 4 per group). (K) Serum E level in male OXGR1OE AG mice treated with AKG for 12 weeks (n= 8 per group). (L-O). Oxygen consumption (L-M) and RER (N-O) of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 8 per group). (P-Q). Representative images (P) and quantification (Q) of gWAT and iWAT HE staining from male OXGR1OE AG mice treated with AKG for 12 weeks (n = 6 per group). (R-S). Representative images (R) and quantification (S) of p-HSL DAB staining from male OXGR1OE AG mice treated with AKG for 12 weeks (n = 6 per group). Results are presented as mean ± SEM. In (A), ** p≤0.01 by non-paired Student’s t test. In (B-C), *p≤0.05, **p≤0.01 by two-way ANOVA followed by post hoc Bonferroni tests. In (E-G), (I-K), (M), (O), (Q) and (S), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Turkey’s tests.
    Figure Legend Snippet: Adrenal specific overexpression of OXGR1 enhances stimulatory effects of AKG on thermogenesis and lipolysis (A). The validation of OXGR1 overexpression. The mRNA expression of OXGR1 was determined in the adrenal glands from male WT control, WT injected with HBAAV2/9-GFP, and WT injected with HBAAV2/9-OXGR1 (OXGR1OE AG ) mice (n=5 per group). (B-C). Body weight gain (B) and cumulative food intake (C) of OXGR1OE AG . Male C57BL/6 mice (8 weeks) were adrenal-specifically injected with control HBAAV2/9-GFP or HBAAV2/9-OXGR1. Two weeks after injections, mice were switched to HFD and further divided into two groups, receiving tap water or water supplemented with 2% AKG for 12 weeks (n = 8 per group). (D-E). Representative image of body composition (D) and fat and lean mass index (E) of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 8 per group). (F-G). Weight index of gWAT (F) and iWAT (G) in male OXGR1OE AG mice treated with AKG for 12 weeks (n = 6 per group). (H-I). Immunoblots (H) and quantification (I) of p-HSL and ATGL protein in the gWAT of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 4 per group). (J). Immunoblots and quantification of UCP1 protein in the BAT of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 4 per group). (K) Serum E level in male OXGR1OE AG mice treated with AKG for 12 weeks (n= 8 per group). (L-O). Oxygen consumption (L-M) and RER (N-O) of male OXGR1OE AG mice treated with AKG for 12 weeks (n = 8 per group). (P-Q). Representative images (P) and quantification (Q) of gWAT and iWAT HE staining from male OXGR1OE AG mice treated with AKG for 12 weeks (n = 6 per group). (R-S). Representative images (R) and quantification (S) of p-HSL DAB staining from male OXGR1OE AG mice treated with AKG for 12 weeks (n = 6 per group). Results are presented as mean ± SEM. In (A), ** p≤0.01 by non-paired Student’s t test. In (B-C), *p≤0.05, **p≤0.01 by two-way ANOVA followed by post hoc Bonferroni tests. In (E-G), (I-K), (M), (O), (Q) and (S), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Turkey’s tests.

    Techniques Used: Over Expression, Expressing, Injection, Mouse Assay, Western Blot, Staining

    OXGR1 is required for metabolic beneficial effects of resistance exercise (A). Body weight gain in male WT littermates and OXGR1KO mice. At 8 weeks of age, male C57BL/6 WT control or OXGR1KO mice were switched to HFD. After 12 weeks of HFD feeding, mice were further divided into two groups, receiving non-exercise or resistance exercise for 14 days. (n = 8 per group). (B). Exercise-induced body weight loss in male WT littermates and OXGR1KO mice. Body weights from exercise mice were subtracted by the average body weight of non-exercise control group for each genotype (n = 8 per group). (C). Exercise-induced fat mass loss in male WT littermates and OXGR1KO mice. Fat mass from exercise mice were subtracted by the average fat mass of non-exercise control group for each genotype (n = 8 per group). (D). Cumulative food intake of male WT littermates and OXGR1KO mice after 14-day resistance exercise (n = 8 per group). (E-F). Weight index of gWAT (E) and iWAT (F) of male OXGR1KO mice after 14-days resistance exercise (n = 8 per group). (G). Body composition of male OXGR1KO mice after 14-days resistance exercise (n = 8 per group). (H). Serum AKG levels of male OXGR1KO mice after resistance exercise. Male OXGR1KO mice (10 weeks) fed with normal chow were receiving resistance exercise for 40 min (n = 8 per group). The serum AKG levels were tested before and immediately after exercise. (I). Serum E level in male OXGR1KO mice after 14-day resistance exercise (n = 8 per group). (J-K). The mRNA expression of UCP1 (J) in the BAT or HSL and ATGL (K) in the gWAT of male OXGR1KO mice after 14-day resistance exercise (n = 4 per group). (L-O). Oxygen consumption (L-M) and RER (N-O) in male OXGR1KO mice after 14-day resistance exercise (n = 8 per group). Results are presented as mean ± SEM. In (A-D) *p≤0.05, ** p≤0.01 by two-way ANOVA followed by post hoc Bonferroni tests. In (H), *p≤0.05 by non-paired Student’s t test. In (E-G), (I-K), (M) and (O), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Tukey’s tests.
    Figure Legend Snippet: OXGR1 is required for metabolic beneficial effects of resistance exercise (A). Body weight gain in male WT littermates and OXGR1KO mice. At 8 weeks of age, male C57BL/6 WT control or OXGR1KO mice were switched to HFD. After 12 weeks of HFD feeding, mice were further divided into two groups, receiving non-exercise or resistance exercise for 14 days. (n = 8 per group). (B). Exercise-induced body weight loss in male WT littermates and OXGR1KO mice. Body weights from exercise mice were subtracted by the average body weight of non-exercise control group for each genotype (n = 8 per group). (C). Exercise-induced fat mass loss in male WT littermates and OXGR1KO mice. Fat mass from exercise mice were subtracted by the average fat mass of non-exercise control group for each genotype (n = 8 per group). (D). Cumulative food intake of male WT littermates and OXGR1KO mice after 14-day resistance exercise (n = 8 per group). (E-F). Weight index of gWAT (E) and iWAT (F) of male OXGR1KO mice after 14-days resistance exercise (n = 8 per group). (G). Body composition of male OXGR1KO mice after 14-days resistance exercise (n = 8 per group). (H). Serum AKG levels of male OXGR1KO mice after resistance exercise. Male OXGR1KO mice (10 weeks) fed with normal chow were receiving resistance exercise for 40 min (n = 8 per group). The serum AKG levels were tested before and immediately after exercise. (I). Serum E level in male OXGR1KO mice after 14-day resistance exercise (n = 8 per group). (J-K). The mRNA expression of UCP1 (J) in the BAT or HSL and ATGL (K) in the gWAT of male OXGR1KO mice after 14-day resistance exercise (n = 4 per group). (L-O). Oxygen consumption (L-M) and RER (N-O) in male OXGR1KO mice after 14-day resistance exercise (n = 8 per group). Results are presented as mean ± SEM. In (A-D) *p≤0.05, ** p≤0.01 by two-way ANOVA followed by post hoc Bonferroni tests. In (H), *p≤0.05 by non-paired Student’s t test. In (E-G), (I-K), (M) and (O), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Tukey’s tests.

    Techniques Used: Mouse Assay, Expressing

    AKG increases fat thermogenesis and lipolysis (A-D). Oxygen consumption (A-B) and respiratory exchange ratio (RER, C-D) in male C57BL/6 mice after 11 weeks of AKG supplementation (n = 8 per group). (E). Body temperature of male mice after 11 weeks of AKG supplementation (n = 9 per group). (F-G). Representative images (F) and quantification (G) of BAT thermogenesis induced by 6 hr cold exposure at 4℃ in male mice supplemented with AKG for 11 weeks (n = 9 per group). (H). The mRNA expression of PPARγ, FASN and ACC in the gWAT and iWAT from male mice supplemented with AKG for 11 weeks (n = 6 per group). (I-J). Immunoblots (I) and quantification (J) of p-HSL and ATGL protein in gWAT of male mice after 11 weeks of AKG supplementation (n = 3 per group). (K-L). DAB staining (K) and quantification (L) of p-HSL in gWAT and iWAT of male mice after 11 weeks of AKG supplementation (n = 9 per group). (M–O). The mRNA expression of thermogenic genes (M) and DAB staining (N) and quantification (O) of UCP1 in BAT of male mice supplemented with AKG for 11 weeks (n = 6-8 per group). (P). The mRNA expression of CD137, CD40, TBX1, TMEM26, CITED1 and slc27a1 in iWAT of male mice supplemented with AKG for 11 weeks (n = 8 per group). (Q–U). Serum levels of NEFA (Q), E (R), NE (S), T3 (T), and T4 (U) in male mice supplemented with AKG for 11 weeks (n = 8-9 per group). Results are presented as mean ± SEM. In (B), (D-E), (G-H), (J), (L-M), and (O-U), *p≤0.05, **p≤0.01, and ***p≤0.001 by non-paired Student’s t test.
    Figure Legend Snippet: AKG increases fat thermogenesis and lipolysis (A-D). Oxygen consumption (A-B) and respiratory exchange ratio (RER, C-D) in male C57BL/6 mice after 11 weeks of AKG supplementation (n = 8 per group). (E). Body temperature of male mice after 11 weeks of AKG supplementation (n = 9 per group). (F-G). Representative images (F) and quantification (G) of BAT thermogenesis induced by 6 hr cold exposure at 4℃ in male mice supplemented with AKG for 11 weeks (n = 9 per group). (H). The mRNA expression of PPARγ, FASN and ACC in the gWAT and iWAT from male mice supplemented with AKG for 11 weeks (n = 6 per group). (I-J). Immunoblots (I) and quantification (J) of p-HSL and ATGL protein in gWAT of male mice after 11 weeks of AKG supplementation (n = 3 per group). (K-L). DAB staining (K) and quantification (L) of p-HSL in gWAT and iWAT of male mice after 11 weeks of AKG supplementation (n = 9 per group). (M–O). The mRNA expression of thermogenic genes (M) and DAB staining (N) and quantification (O) of UCP1 in BAT of male mice supplemented with AKG for 11 weeks (n = 6-8 per group). (P). The mRNA expression of CD137, CD40, TBX1, TMEM26, CITED1 and slc27a1 in iWAT of male mice supplemented with AKG for 11 weeks (n = 8 per group). (Q–U). Serum levels of NEFA (Q), E (R), NE (S), T3 (T), and T4 (U) in male mice supplemented with AKG for 11 weeks (n = 8-9 per group). Results are presented as mean ± SEM. In (B), (D-E), (G-H), (J), (L-M), and (O-U), *p≤0.05, **p≤0.01, and ***p≤0.001 by non-paired Student’s t test.

    Techniques Used: Mouse Assay, Expressing, Western Blot, Staining

    Adrenal specific reexpression of OXGR1 rescues the stimulatory effects of AKG on thermogenesis and lipolysis (A). Serum E level in male OXGR1KO mice. At 12 weeks of age, male control or OXGR1KO mice were switched to HFD and received tap water or water supplemented with 2% AKG for 13 weeks (n = 8 per group). (B). Immunoblots and quantification of UCP1 protein expression in the BAT of male OXGR1KO mice treated with AKG for 13 weeks (n = 4 per group). (C-D). Representative images (C) and quantification (D) of iWAT and gWAT HE staining from male OXGR1KO mice treated with AKG for 13 weeks (n = 6 per group). (E-F). Representative images (E) and quantification (F) of p-HSL DAB staining from male OXGR1KO mice treated with AKG for 13 weeks (n = 6 per group). (G). The validation of OXGR1 reexpression. The mRNA expression of OXGR1 was determined in the adrenal glands from male WT control, OXGR1KO injected with HBAAV2/9-GFP, and OXGR1KO injected with HBAAV2/9-OXGR1 (OXGR1RE AG ) mice. (H). Serum E level in male OXGR1RE AG . Male OXGR1KO mice (8 weeks) were adrenal-specifically injected with control HBAAV2/9-GFP or HBAAV2/9-OXGR1. Two weeks after injections, mice were switched to HFD and further divided into two groups, receiving tap water or 2% AKG for 13 weeks. (n = 6 per group). (I). Immunoblots and quantification of UCP1 protein expression in the BAT of OXGR1RE AG mice treated with AKG for 13 weeks (n = 4 per group). (J-K). Representative images (J) and quantification (K) of iWAT and gWAT HE staining from OXGR1RE AG mice treated with AKG for 13 weeks (n = 6 per group). (L-M). Representative images (L) and quantification (M) of p-HSL DAB staining from OXGR1RE AG mice treated with AKG for 13 weeks (n = 6 per group). Results are presented as mean ± SEM. In (A-B), (D), (F), (H-I), (K) and (M), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Turkey’s tests.
    Figure Legend Snippet: Adrenal specific reexpression of OXGR1 rescues the stimulatory effects of AKG on thermogenesis and lipolysis (A). Serum E level in male OXGR1KO mice. At 12 weeks of age, male control or OXGR1KO mice were switched to HFD and received tap water or water supplemented with 2% AKG for 13 weeks (n = 8 per group). (B). Immunoblots and quantification of UCP1 protein expression in the BAT of male OXGR1KO mice treated with AKG for 13 weeks (n = 4 per group). (C-D). Representative images (C) and quantification (D) of iWAT and gWAT HE staining from male OXGR1KO mice treated with AKG for 13 weeks (n = 6 per group). (E-F). Representative images (E) and quantification (F) of p-HSL DAB staining from male OXGR1KO mice treated with AKG for 13 weeks (n = 6 per group). (G). The validation of OXGR1 reexpression. The mRNA expression of OXGR1 was determined in the adrenal glands from male WT control, OXGR1KO injected with HBAAV2/9-GFP, and OXGR1KO injected with HBAAV2/9-OXGR1 (OXGR1RE AG ) mice. (H). Serum E level in male OXGR1RE AG . Male OXGR1KO mice (8 weeks) were adrenal-specifically injected with control HBAAV2/9-GFP or HBAAV2/9-OXGR1. Two weeks after injections, mice were switched to HFD and further divided into two groups, receiving tap water or 2% AKG for 13 weeks. (n = 6 per group). (I). Immunoblots and quantification of UCP1 protein expression in the BAT of OXGR1RE AG mice treated with AKG for 13 weeks (n = 4 per group). (J-K). Representative images (J) and quantification (K) of iWAT and gWAT HE staining from OXGR1RE AG mice treated with AKG for 13 weeks (n = 6 per group). (L-M). Representative images (L) and quantification (M) of p-HSL DAB staining from OXGR1RE AG mice treated with AKG for 13 weeks (n = 6 per group). Results are presented as mean ± SEM. In (A-B), (D), (F), (H-I), (K) and (M), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Turkey’s tests.

    Techniques Used: Mouse Assay, Western Blot, Expressing, Staining, Injection

    Metabolic effects of AKG is mediated by adrenergic stimulation of thermogenesis and lipolysis (A). Serum AKG concentration-time profile obtained from male C57BL/6 mice (10 weeks) fed with normal chow before or after i.p AKG (10 mg/kg body weight). The serum AKG level were tested at 0, 1, 2, 4 and 6 hrs after injection (n = 8 per group). (B-C). Representative images (B) and quantification (C) of BAT thermogenesis after 6 hr cold exposure at 4℃. Male C57BL/6 mice (10 weeks) were i.p. injected with 10 mg/kg AKG or saline and immediately exposed to cold stress at 4℃ (n = 8 per group). (D). Immunoblots and quantification of p-HSL and ATGL in the gWAT of male C57BL/6 mice (10 weeks) 3 hrs after i.p. injection of 10 mg/kg AKG or saline (n = 3 per group). (E). Serum E level in AKG treated male mice 3 hrs after i.p. injection (n = 8 per group). (F-I). Oxygen consumption (F-G) and RER (H-I) in male C57BL/6 mice (10 weeks) i.p. injected with saline, 10 mg/kg AKG, 1 mg/kg SR59230A (ADRB3 inhibitor) or AKG + SR59230A (n = 8 per group). All injections were performed at 7:00 am of second day. Data was summarized in bar graph (G and I) by light or dark cycle of second day. (J-N). Body weight gain (J), cumulative food intake (K), body composition (L), fat weight (M) and serum NEFA (N) of shame or adrenalectomized male C57BL/6 mice. Male mice were adrenalectomized at 8 weeks of age. Two weeks after surgeries, male mice were switched to HFD and given free access to tap water or 2% AKG for 9 weeks (n = 8 per group). (O-P). Representative images (O) and quantification (P) of BAT thermogenesis after 6h cold exposure at 4℃ in shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 8 per group). (Q). The mRNA expression of themogenic genes in the BAT of shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 6 per group). (R-S) Immunoblots (R) and quantification (S) of p-HSL and ATGL protein in the gWAT of shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 4 per group). (T-U). Immunoblots (T) and quantification (U) of UCP1 protein in the BAT of shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 4 per group). Results are presented as mean ± SEM. In (A), *p≤0.05 by non-paired Student’s t test compared with before injection. In (C-E), *p≤0.05, **p≤ 0.01 by non-paired Student’s t test. In (J-K), *p≤0.05 by two-way ANOVA followed by post hoc Bonferroni tests. In (G), (I), (L-N), (P-Q), (S) and (U), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Tukey’s tests.
    Figure Legend Snippet: Metabolic effects of AKG is mediated by adrenergic stimulation of thermogenesis and lipolysis (A). Serum AKG concentration-time profile obtained from male C57BL/6 mice (10 weeks) fed with normal chow before or after i.p AKG (10 mg/kg body weight). The serum AKG level were tested at 0, 1, 2, 4 and 6 hrs after injection (n = 8 per group). (B-C). Representative images (B) and quantification (C) of BAT thermogenesis after 6 hr cold exposure at 4℃. Male C57BL/6 mice (10 weeks) were i.p. injected with 10 mg/kg AKG or saline and immediately exposed to cold stress at 4℃ (n = 8 per group). (D). Immunoblots and quantification of p-HSL and ATGL in the gWAT of male C57BL/6 mice (10 weeks) 3 hrs after i.p. injection of 10 mg/kg AKG or saline (n = 3 per group). (E). Serum E level in AKG treated male mice 3 hrs after i.p. injection (n = 8 per group). (F-I). Oxygen consumption (F-G) and RER (H-I) in male C57BL/6 mice (10 weeks) i.p. injected with saline, 10 mg/kg AKG, 1 mg/kg SR59230A (ADRB3 inhibitor) or AKG + SR59230A (n = 8 per group). All injections were performed at 7:00 am of second day. Data was summarized in bar graph (G and I) by light or dark cycle of second day. (J-N). Body weight gain (J), cumulative food intake (K), body composition (L), fat weight (M) and serum NEFA (N) of shame or adrenalectomized male C57BL/6 mice. Male mice were adrenalectomized at 8 weeks of age. Two weeks after surgeries, male mice were switched to HFD and given free access to tap water or 2% AKG for 9 weeks (n = 8 per group). (O-P). Representative images (O) and quantification (P) of BAT thermogenesis after 6h cold exposure at 4℃ in shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 8 per group). (Q). The mRNA expression of themogenic genes in the BAT of shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 6 per group). (R-S) Immunoblots (R) and quantification (S) of p-HSL and ATGL protein in the gWAT of shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 4 per group). (T-U). Immunoblots (T) and quantification (U) of UCP1 protein in the BAT of shame or adrenalectomized male mice treated with AKG for 9 weeks (n = 4 per group). Results are presented as mean ± SEM. In (A), *p≤0.05 by non-paired Student’s t test compared with before injection. In (C-E), *p≤0.05, **p≤ 0.01 by non-paired Student’s t test. In (J-K), *p≤0.05 by two-way ANOVA followed by post hoc Bonferroni tests. In (G), (I), (L-N), (P-Q), (S) and (U), different letters between bars indicate p≤0.05 by one-way ANOVA followed by post hoc Tukey’s tests.

    Techniques Used: Concentration Assay, Mouse Assay, Injection, Western Blot, Expressing

    18) Product Images from "UDP-Glucuronosyltransferase 1a Enzymes Are Present and Active in the Mouse Blastocyst"

    Article Title: UDP-Glucuronosyltransferase 1a Enzymes Are Present and Active in the Mouse Blastocyst

    Journal: Drug Metabolism and Disposition

    doi: 10.1124/dmd.114.059766

    Detection of Ugt proteins in mouse blastocysts. (A) Proteins of the Ugt1a family were present as detected by a UGT1A/Ugt1a pan-specific antibody (55 kDa). (B) Murine Ugt1a1 was not detected (52 kDa). (C) Murine Ugt1a3 was not detected (37 kDa). (D) Murine
    Figure Legend Snippet: Detection of Ugt proteins in mouse blastocysts. (A) Proteins of the Ugt1a family were present as detected by a UGT1A/Ugt1a pan-specific antibody (55 kDa). (B) Murine Ugt1a1 was not detected (52 kDa). (C) Murine Ugt1a3 was not detected (37 kDa). (D) Murine

    Techniques Used:

    Confocal immunofluorescence analysis of Ugt expression and localization in blastocysts. (A) Exemplary images of blastocysts stained with pan-specific antibodies against Ugt1a and Ugt2b, with DAPI staining to show the cell nuclei. Strong Ugt1a signal is
    Figure Legend Snippet: Confocal immunofluorescence analysis of Ugt expression and localization in blastocysts. (A) Exemplary images of blastocysts stained with pan-specific antibodies against Ugt1a and Ugt2b, with DAPI staining to show the cell nuclei. Strong Ugt1a signal is

    Techniques Used: Immunofluorescence, Expressing, Staining

    19) Product Images from "Cis and Trans Regulatory Mechanisms Control AP2-Mediated B Cell Receptor Endocytosis via Select Tyrosine-Based Motifs"

    Article Title: Cis and Trans Regulatory Mechanisms Control AP2-Mediated B Cell Receptor Endocytosis via Select Tyrosine-Based Motifs

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0054938

    Ultrastructural Colocalization of Ligand-BCR Complexes and AP2 in Clathrin-Coated Pits. A20µWT B cells or primary murine splenocytes were pulsed with anti-IgM-btn followed by anti-biotin-15 nm gold (arrow heads), incubated 2 minutes at 37° and then plasma membrane rips were prepared as previously reported [10] . The exposed cytoplasmic face of the plasma membrane was stained with anti-AP2 and Protein A-5 nm gold (arrows). The percent of BCR-containing CCP that also stained for AP2 is indicated below each image. Inset: AP2 and BCR co-localization within electron dense membrane regions lacking discernable CCP architecture. Shown are representative images from 1 of 3 experiments, with 1,000+ BCR-bound gold particles or 100+ CCP photographed cumulatively.
    Figure Legend Snippet: Ultrastructural Colocalization of Ligand-BCR Complexes and AP2 in Clathrin-Coated Pits. A20µWT B cells or primary murine splenocytes were pulsed with anti-IgM-btn followed by anti-biotin-15 nm gold (arrow heads), incubated 2 minutes at 37° and then plasma membrane rips were prepared as previously reported [10] . The exposed cytoplasmic face of the plasma membrane was stained with anti-AP2 and Protein A-5 nm gold (arrows). The percent of BCR-containing CCP that also stained for AP2 is indicated below each image. Inset: AP2 and BCR co-localization within electron dense membrane regions lacking discernable CCP architecture. Shown are representative images from 1 of 3 experiments, with 1,000+ BCR-bound gold particles or 100+ CCP photographed cumulatively.

    Techniques Used: Incubation, Staining

    20) Product Images from "Flavivirus Infection Activates the XBP1 Pathway of the Unfolded Protein Response To Cope with Endoplasmic Reticulum Stress ▿"

    Article Title: Flavivirus Infection Activates the XBP1 Pathway of the Unfolded Protein Response To Cope with Endoplasmic Reticulum Stress ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.00879-06

    Flavivirus induces XBP1 mRNA splicing in infected cultured neuronal cells and mouse brains. (A) The analysis scheme of XBP1 mRNA splicing. The relative locations of the 26-nt intron, the PstI recognition site, and the PCR-amplified region are shown. The sizes of PCR-amplified fragments from spliced and unspliced XBP1 with or without PstI cleavage are also listed. (B) XBP1 mRNA splicing and induction of downstream genes. N18, a mouse neuroblastoma cell line, was mock infected (lanes 1 to 4), treated with tunicamycin (1 μg/ml; lanes 5 to 8), and infected with JEV (lanes 9 to 12) or DEN-2 (lanes 13 to 16). The MOI was 3. Cells were harvested at 6, 12, 24, or 36 h after treatment for RT-PCR analysis using primer sets specific for the genes shown on the left. The PCR product of XBP1 was further analyzed by PstI digestion. DNA molecules were separated by 2 or 1.5% agarose gel electrophoresis, stained with ethidium bromide, and photographed with a Photo-Print photodocumentation system (Vilber Lourmat). (C) The induction ( n -fold) of selected UPR downstream genes was quantified by real-time RT-PCR as described in Materials and Methods. (D) Decrease in full-length ATF6 (p90ATF6) in flavivirus-infected cells. N18 cells were treated with 2 mM dithiothreitol (DTT) for 3 h or infected with JEV or DEN-2 (MOI, 5) for 20 h before the cell lysates were harvested (62.5 mM Tris-HCl, pH 6.8, 2% SDS, 10% glycerol) for immunoblotting with anti-ATF6 (IMGENEX) and anti-actin antibodies. (E) The protein expression of XBP1s in N18 cells was analyzed by immunoblotting at several time points after infection with JEV or DEN-2 (MOI, 5) as indicated. The nuclear extracts were separated by 10% SDS-polyacrylamide gel electrophoresis and immunoblotted with the antibody indicated on the right. (F) The XBP1 mRNA status in brain lysates of C57/B6 mice intracranially inoculated with JEV (1 × 10 4 PFU/mouse) was analyzed by RT-PCR and PstI digestion. The level of JEV replication was also detected by RT-PCR of the NS3 region of the JEV genome. PBS, mice sham inoculated with PBS serving as a negative control. p.i., postinfection.
    Figure Legend Snippet: Flavivirus induces XBP1 mRNA splicing in infected cultured neuronal cells and mouse brains. (A) The analysis scheme of XBP1 mRNA splicing. The relative locations of the 26-nt intron, the PstI recognition site, and the PCR-amplified region are shown. The sizes of PCR-amplified fragments from spliced and unspliced XBP1 with or without PstI cleavage are also listed. (B) XBP1 mRNA splicing and induction of downstream genes. N18, a mouse neuroblastoma cell line, was mock infected (lanes 1 to 4), treated with tunicamycin (1 μg/ml; lanes 5 to 8), and infected with JEV (lanes 9 to 12) or DEN-2 (lanes 13 to 16). The MOI was 3. Cells were harvested at 6, 12, 24, or 36 h after treatment for RT-PCR analysis using primer sets specific for the genes shown on the left. The PCR product of XBP1 was further analyzed by PstI digestion. DNA molecules were separated by 2 or 1.5% agarose gel electrophoresis, stained with ethidium bromide, and photographed with a Photo-Print photodocumentation system (Vilber Lourmat). (C) The induction ( n -fold) of selected UPR downstream genes was quantified by real-time RT-PCR as described in Materials and Methods. (D) Decrease in full-length ATF6 (p90ATF6) in flavivirus-infected cells. N18 cells were treated with 2 mM dithiothreitol (DTT) for 3 h or infected with JEV or DEN-2 (MOI, 5) for 20 h before the cell lysates were harvested (62.5 mM Tris-HCl, pH 6.8, 2% SDS, 10% glycerol) for immunoblotting with anti-ATF6 (IMGENEX) and anti-actin antibodies. (E) The protein expression of XBP1s in N18 cells was analyzed by immunoblotting at several time points after infection with JEV or DEN-2 (MOI, 5) as indicated. The nuclear extracts were separated by 10% SDS-polyacrylamide gel electrophoresis and immunoblotted with the antibody indicated on the right. (F) The XBP1 mRNA status in brain lysates of C57/B6 mice intracranially inoculated with JEV (1 × 10 4 PFU/mouse) was analyzed by RT-PCR and PstI digestion. The level of JEV replication was also detected by RT-PCR of the NS3 region of the JEV genome. PBS, mice sham inoculated with PBS serving as a negative control. p.i., postinfection.

    Techniques Used: Infection, Cell Culture, Polymerase Chain Reaction, Amplification, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining, Quantitative RT-PCR, Expressing, Polyacrylamide Gel Electrophoresis, Mouse Assay, Negative Control

    21) Product Images from "Characterization of novel biomarkers in selecting for subtype specific medulloblastoma phenotypes"

    Article Title: Characterization of novel biomarkers in selecting for subtype specific medulloblastoma phenotypes

    Journal: Oncotarget

    doi:

    Candidate cell surface markers are differentially expressed in MB cell lines/primary cultures and patient samples A. Representative dot plots of staining for candidate biomarkers in Daoy, MED311-FH and UI226 tumorspheres (SHH variant) vs . tumorspheres from Group 3/Group 4 cell lines by flow cytometry. Insets: respective isotype controls. 7AAD: 7-aminoactinomycin D cell viability dye. B. - D. CD271 (B), CD171 (C) and CD106 (D) expression in paraffin embedded sections of fetal cerebellum and primary medulloblastoma samples. Scale bar: 400 μm.
    Figure Legend Snippet: Candidate cell surface markers are differentially expressed in MB cell lines/primary cultures and patient samples A. Representative dot plots of staining for candidate biomarkers in Daoy, MED311-FH and UI226 tumorspheres (SHH variant) vs . tumorspheres from Group 3/Group 4 cell lines by flow cytometry. Insets: respective isotype controls. 7AAD: 7-aminoactinomycin D cell viability dye. B. - D. CD271 (B), CD171 (C) and CD106 (D) expression in paraffin embedded sections of fetal cerebellum and primary medulloblastoma samples. Scale bar: 400 μm.

    Techniques Used: Staining, Variant Assay, Flow Cytometry, Cytometry, Expressing

    Transcript levels of CD271/p75NTR, CD171L1CAM, EGFR and CD106/VCAM1 across the 4 MB molecular variants Note that the CD271 Toronto dataset was previously published in Neoplasia, 15, Morrison et al., Deconstruction of Medulloblastoma cellular heterogeneity reveals differences between the most highly invasion and self-renewing phenotypes , 384-398, Copyright Elsevier (2013). A. - C. Gene expression data from 3 independent transcriptome datasets representing 548 patient samples showing downregulation of CD171/NCAM-L1 and relative enrichment of CD271/p75NTR, CD106/VCAM1, and EGFR, in SHH tumors compared with the other variants. Bars denote 1.5 interquartile range within each group. All subgroups were compared using a Kruskal-Wallis test for significance. Data are presented as log2 -transformed signal intensity. P
    Figure Legend Snippet: Transcript levels of CD271/p75NTR, CD171L1CAM, EGFR and CD106/VCAM1 across the 4 MB molecular variants Note that the CD271 Toronto dataset was previously published in Neoplasia, 15, Morrison et al., Deconstruction of Medulloblastoma cellular heterogeneity reveals differences between the most highly invasion and self-renewing phenotypes , 384-398, Copyright Elsevier (2013). A. - C. Gene expression data from 3 independent transcriptome datasets representing 548 patient samples showing downregulation of CD171/NCAM-L1 and relative enrichment of CD271/p75NTR, CD106/VCAM1, and EGFR, in SHH tumors compared with the other variants. Bars denote 1.5 interquartile range within each group. All subgroups were compared using a Kruskal-Wallis test for significance. Data are presented as log2 -transformed signal intensity. P

    Techniques Used: Expressing, Transformation Assay

    22) Product Images from "Characterization of novel biomarkers in selecting for subtype specific medulloblastoma phenotypes"

    Article Title: Characterization of novel biomarkers in selecting for subtype specific medulloblastoma phenotypes

    Journal: Oncotarget

    doi:

    Candidate cell surface markers are differentially expressed in MB cell lines/primary cultures and patient samples A. Representative dot plots of staining for candidate biomarkers in Daoy, MED311-FH and UI226 tumorspheres (SHH variant) vs . tumorspheres from Group 3/Group 4 cell lines by flow cytometry. Insets: respective isotype controls. 7AAD: 7-aminoactinomycin D cell viability dye. B. - D. CD271 (B), CD171 (C) and CD106 (D) expression in paraffin embedded sections of fetal cerebellum and primary medulloblastoma samples. Scale bar: 400 μm.
    Figure Legend Snippet: Candidate cell surface markers are differentially expressed in MB cell lines/primary cultures and patient samples A. Representative dot plots of staining for candidate biomarkers in Daoy, MED311-FH and UI226 tumorspheres (SHH variant) vs . tumorspheres from Group 3/Group 4 cell lines by flow cytometry. Insets: respective isotype controls. 7AAD: 7-aminoactinomycin D cell viability dye. B. - D. CD271 (B), CD171 (C) and CD106 (D) expression in paraffin embedded sections of fetal cerebellum and primary medulloblastoma samples. Scale bar: 400 μm.

    Techniques Used: Staining, Variant Assay, Flow Cytometry, Cytometry, Expressing

    Transcript levels of CD271/p75NTR, CD171L1CAM, EGFR and CD106/VCAM1 across the 4 MB molecular variants Note that the CD271 Toronto dataset was previously published in Neoplasia, 15, Morrison et al., Deconstruction of Medulloblastoma cellular heterogeneity reveals differences between the most highly invasion and self-renewing phenotypes , 384-398, Copyright Elsevier (2013). A. - C. Gene expression data from 3 independent transcriptome datasets representing 548 patient samples showing downregulation of CD171/NCAM-L1 and relative enrichment of CD271/p75NTR, CD106/VCAM1, and EGFR, in SHH tumors compared with the other variants. Bars denote 1.5 interquartile range within each group. All subgroups were compared using a Kruskal-Wallis test for significance. Data are presented as log2 -transformed signal intensity. P
    Figure Legend Snippet: Transcript levels of CD271/p75NTR, CD171L1CAM, EGFR and CD106/VCAM1 across the 4 MB molecular variants Note that the CD271 Toronto dataset was previously published in Neoplasia, 15, Morrison et al., Deconstruction of Medulloblastoma cellular heterogeneity reveals differences between the most highly invasion and self-renewing phenotypes , 384-398, Copyright Elsevier (2013). A. - C. Gene expression data from 3 independent transcriptome datasets representing 548 patient samples showing downregulation of CD171/NCAM-L1 and relative enrichment of CD271/p75NTR, CD106/VCAM1, and EGFR, in SHH tumors compared with the other variants. Bars denote 1.5 interquartile range within each group. All subgroups were compared using a Kruskal-Wallis test for significance. Data are presented as log2 -transformed signal intensity. P

    Techniques Used: Expressing, Transformation Assay

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    SDS Page:

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    Jackson Immuno cd106
    Candidate cell surface markers are differentially expressed in MB cell lines/primary cultures and patient samples A. Representative dot plots of staining for candidate biomarkers in Daoy, MED311-FH and UI226 tumorspheres (SHH variant) vs . tumorspheres from Group 3/Group 4 cell lines by flow cytometry. Insets: respective isotype controls. 7AAD: 7-aminoactinomycin D cell viability dye. B. - D. CD271 (B), CD171 (C) and <t>CD106</t> (D) expression in paraffin embedded sections of fetal cerebellum and primary medulloblastoma samples. Scale bar: 400 μm.
    Cd106, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd106/product/Jackson Immuno
    Average 99 stars, based on 1 article reviews
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    Jackson Immuno biotin sp conjugated affinipure goat anti rabbit immunoglobulin g
    Candidate cell surface markers are differentially expressed in MB cell lines/primary cultures and patient samples A. Representative dot plots of staining for candidate biomarkers in Daoy, MED311-FH and UI226 tumorspheres (SHH variant) vs . tumorspheres from Group 3/Group 4 cell lines by flow cytometry. Insets: respective isotype controls. 7AAD: 7-aminoactinomycin D cell viability dye. B. - D. CD271 (B), CD171 (C) and <t>CD106</t> (D) expression in paraffin embedded sections of fetal cerebellum and primary medulloblastoma samples. Scale bar: 400 μm.
    Biotin Sp Conjugated Affinipure Goat Anti Rabbit Immunoglobulin G, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotin sp conjugated affinipure goat anti rabbit immunoglobulin g/product/Jackson Immuno
    Average 85 stars, based on 6 article reviews
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    Jackson Immuno biotin sp affinipure goat anti rabbit mouse immunoglobulin g
    Candidate cell surface markers are differentially expressed in MB cell lines/primary cultures and patient samples A. Representative dot plots of staining for candidate biomarkers in Daoy, MED311-FH and UI226 tumorspheres (SHH variant) vs . tumorspheres from Group 3/Group 4 cell lines by flow cytometry. Insets: respective isotype controls. 7AAD: 7-aminoactinomycin D cell viability dye. B. - D. CD271 (B), CD171 (C) and <t>CD106</t> (D) expression in paraffin embedded sections of fetal cerebellum and primary medulloblastoma samples. Scale bar: 400 μm.
    Biotin Sp Affinipure Goat Anti Rabbit Mouse Immunoglobulin G, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotin sp affinipure goat anti rabbit mouse immunoglobulin g/product/Jackson Immuno
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    biotin sp affinipure goat anti rabbit mouse immunoglobulin g - by Bioz Stars, 2020-09
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    Candidate cell surface markers are differentially expressed in MB cell lines/primary cultures and patient samples A. Representative dot plots of staining for candidate biomarkers in Daoy, MED311-FH and UI226 tumorspheres (SHH variant) vs . tumorspheres from Group 3/Group 4 cell lines by flow cytometry. Insets: respective isotype controls. 7AAD: 7-aminoactinomycin D cell viability dye. B. - D. CD271 (B), CD171 (C) and CD106 (D) expression in paraffin embedded sections of fetal cerebellum and primary medulloblastoma samples. Scale bar: 400 μm.

    Journal: Oncotarget

    Article Title: Characterization of novel biomarkers in selecting for subtype specific medulloblastoma phenotypes

    doi:

    Figure Lengend Snippet: Candidate cell surface markers are differentially expressed in MB cell lines/primary cultures and patient samples A. Representative dot plots of staining for candidate biomarkers in Daoy, MED311-FH and UI226 tumorspheres (SHH variant) vs . tumorspheres from Group 3/Group 4 cell lines by flow cytometry. Insets: respective isotype controls. 7AAD: 7-aminoactinomycin D cell viability dye. B. - D. CD271 (B), CD171 (C) and CD106 (D) expression in paraffin embedded sections of fetal cerebellum and primary medulloblastoma samples. Scale bar: 400 μm.

    Article Snippet: Slides were washed in (CD271): 1XPBS, (CD171/CD106): 1XTBS and treated with secondary antibody: for CD271 (1:500), for CD106 (1:200) (Biotin-SP-Affinipure Goat Anti-Rabbit IgG (Jackson Immunoresearch, West Grove, PA, USA) and for CD171 (1:200) (Biotin-SP-AffiniPure Sheep Anti-Mouse IgG (H+L) (Jackson Immunoresearch) diluted in (CD271): 1% lamb serum in 1XPBS, (CD171/CD106): 1% goat serum / 1% BSA diluted in TBS for 2 hours at room temperature.

    Techniques: Staining, Variant Assay, Flow Cytometry, Cytometry, Expressing

    Transcript levels of CD271/p75NTR, CD171L1CAM, EGFR and CD106/VCAM1 across the 4 MB molecular variants Note that the CD271 Toronto dataset was previously published in Neoplasia, 15, Morrison et al., Deconstruction of Medulloblastoma cellular heterogeneity reveals differences between the most highly invasion and self-renewing phenotypes , 384-398, Copyright Elsevier (2013). A. - C. Gene expression data from 3 independent transcriptome datasets representing 548 patient samples showing downregulation of CD171/NCAM-L1 and relative enrichment of CD271/p75NTR, CD106/VCAM1, and EGFR, in SHH tumors compared with the other variants. Bars denote 1.5 interquartile range within each group. All subgroups were compared using a Kruskal-Wallis test for significance. Data are presented as log2 -transformed signal intensity. P

    Journal: Oncotarget

    Article Title: Characterization of novel biomarkers in selecting for subtype specific medulloblastoma phenotypes

    doi:

    Figure Lengend Snippet: Transcript levels of CD271/p75NTR, CD171L1CAM, EGFR and CD106/VCAM1 across the 4 MB molecular variants Note that the CD271 Toronto dataset was previously published in Neoplasia, 15, Morrison et al., Deconstruction of Medulloblastoma cellular heterogeneity reveals differences between the most highly invasion and self-renewing phenotypes , 384-398, Copyright Elsevier (2013). A. - C. Gene expression data from 3 independent transcriptome datasets representing 548 patient samples showing downregulation of CD171/NCAM-L1 and relative enrichment of CD271/p75NTR, CD106/VCAM1, and EGFR, in SHH tumors compared with the other variants. Bars denote 1.5 interquartile range within each group. All subgroups were compared using a Kruskal-Wallis test for significance. Data are presented as log2 -transformed signal intensity. P

    Article Snippet: Slides were washed in (CD271): 1XPBS, (CD171/CD106): 1XTBS and treated with secondary antibody: for CD271 (1:500), for CD106 (1:200) (Biotin-SP-Affinipure Goat Anti-Rabbit IgG (Jackson Immunoresearch, West Grove, PA, USA) and for CD171 (1:200) (Biotin-SP-AffiniPure Sheep Anti-Mouse IgG (H+L) (Jackson Immunoresearch) diluted in (CD271): 1% lamb serum in 1XPBS, (CD171/CD106): 1% goat serum / 1% BSA diluted in TBS for 2 hours at room temperature.

    Techniques: Expressing, Transformation Assay