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  • 99
    Qiagen qiagen easy minikit
    Qiagen Easy Minikit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 707 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher qiagen rneasy minikit
    Qiagen Rneasy Minikit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen qiagen rneasy minikit
    si <t>RNA</t> knockdown of TRPM 7 channel abrogates Ca 2+ influx and I spont in <t>K562</t> cells. (A) detection of TRPM 7 transcripts (398 bp) by conventional RT ‐ PCR . Ten μ mol/L doxycycline ( DXC ) almost completely eliminated TRPM 7 mRNA expression. (B) immunoblots of K562 protein extracts by TRPM 7 antibody. Doxycycline concentration dependently (0.1–10 μ mol/L) reduced TRPM 7 protein expression. The results shown in A and B are representative of three independent experiments. (C) fura‐2 Ca 2+ fluorescence imaging from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing tet ‐inducible TRPM 7‐specific si RNA (si TRPM 7). n = 36 and 26, respectively. (D) density of I spont recorded by conventional whole‐cell recording from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing si TRPM 7. To facilitate the induction of TRPM 7 currents, ATP and Mg 2+ were absent in the patch pipette. The numbers of cells tested are shown in parentheses.
    Qiagen Rneasy Minikit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 14454 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen rneasy lipid tissue minikit
    si <t>RNA</t> knockdown of TRPM 7 channel abrogates Ca 2+ influx and I spont in <t>K562</t> cells. (A) detection of TRPM 7 transcripts (398 bp) by conventional RT ‐ PCR . Ten μ mol/L doxycycline ( DXC ) almost completely eliminated TRPM 7 mRNA expression. (B) immunoblots of K562 protein extracts by TRPM 7 antibody. Doxycycline concentration dependently (0.1–10 μ mol/L) reduced TRPM 7 protein expression. The results shown in A and B are representative of three independent experiments. (C) fura‐2 Ca 2+ fluorescence imaging from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing tet ‐inducible TRPM 7‐specific si RNA (si TRPM 7). n = 36 and 26, respectively. (D) density of I spont recorded by conventional whole‐cell recording from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing si TRPM 7. To facilitate the induction of TRPM 7 currents, ATP and Mg 2+ were absent in the patch pipette. The numbers of cells tested are shown in parentheses.
    Rneasy Lipid Tissue Minikit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 321 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen rneasy fibrous tissue minikit
    si <t>RNA</t> knockdown of TRPM 7 channel abrogates Ca 2+ influx and I spont in <t>K562</t> cells. (A) detection of TRPM 7 transcripts (398 bp) by conventional RT ‐ PCR . Ten μ mol/L doxycycline ( DXC ) almost completely eliminated TRPM 7 mRNA expression. (B) immunoblots of K562 protein extracts by TRPM 7 antibody. Doxycycline concentration dependently (0.1–10 μ mol/L) reduced TRPM 7 protein expression. The results shown in A and B are representative of three independent experiments. (C) fura‐2 Ca 2+ fluorescence imaging from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing tet ‐inducible TRPM 7‐specific si RNA (si TRPM 7). n = 36 and 26, respectively. (D) density of I spont recorded by conventional whole‐cell recording from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing si TRPM 7. To facilitate the induction of TRPM 7 currents, ATP and Mg 2+ were absent in the patch pipette. The numbers of cells tested are shown in parentheses.
    Rneasy Fibrous Tissue Minikit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 111 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen rneasy protect cell minikit
    si <t>RNA</t> knockdown of TRPM 7 channel abrogates Ca 2+ influx and I spont in <t>K562</t> cells. (A) detection of TRPM 7 transcripts (398 bp) by conventional RT ‐ PCR . Ten μ mol/L doxycycline ( DXC ) almost completely eliminated TRPM 7 mRNA expression. (B) immunoblots of K562 protein extracts by TRPM 7 antibody. Doxycycline concentration dependently (0.1–10 μ mol/L) reduced TRPM 7 protein expression. The results shown in A and B are representative of three independent experiments. (C) fura‐2 Ca 2+ fluorescence imaging from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing tet ‐inducible TRPM 7‐specific si RNA (si TRPM 7). n = 36 and 26, respectively. (D) density of I spont recorded by conventional whole‐cell recording from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing si TRPM 7. To facilitate the induction of TRPM 7 currents, ATP and Mg 2+ were absent in the patch pipette. The numbers of cells tested are shown in parentheses.
    Rneasy Protect Cell Minikit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen rneasy plus universal minikit
    si <t>RNA</t> knockdown of TRPM 7 channel abrogates Ca 2+ influx and I spont in <t>K562</t> cells. (A) detection of TRPM 7 transcripts (398 bp) by conventional RT ‐ PCR . Ten μ mol/L doxycycline ( DXC ) almost completely eliminated TRPM 7 mRNA expression. (B) immunoblots of K562 protein extracts by TRPM 7 antibody. Doxycycline concentration dependently (0.1–10 μ mol/L) reduced TRPM 7 protein expression. The results shown in A and B are representative of three independent experiments. (C) fura‐2 Ca 2+ fluorescence imaging from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing tet ‐inducible TRPM 7‐specific si RNA (si TRPM 7). n = 36 and 26, respectively. (D) density of I spont recorded by conventional whole‐cell recording from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing si TRPM 7. To facilitate the induction of TRPM 7 currents, ATP and Mg 2+ were absent in the patch pipette. The numbers of cells tested are shown in parentheses.
    Rneasy Plus Universal Minikit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 4778 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen rneasy protect bacteria minikit
    si <t>RNA</t> knockdown of TRPM 7 channel abrogates Ca 2+ influx and I spont in <t>K562</t> cells. (A) detection of TRPM 7 transcripts (398 bp) by conventional RT ‐ PCR . Ten μ mol/L doxycycline ( DXC ) almost completely eliminated TRPM 7 mRNA expression. (B) immunoblots of K562 protein extracts by TRPM 7 antibody. Doxycycline concentration dependently (0.1–10 μ mol/L) reduced TRPM 7 protein expression. The results shown in A and B are representative of three independent experiments. (C) fura‐2 Ca 2+ fluorescence imaging from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing tet ‐inducible TRPM 7‐specific si RNA (si TRPM 7). n = 36 and 26, respectively. (D) density of I spont recorded by conventional whole‐cell recording from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing si TRPM 7. To facilitate the induction of TRPM 7 currents, ATP and Mg 2+ were absent in the patch pipette. The numbers of cells tested are shown in parentheses.
    Rneasy Protect Bacteria Minikit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen rneasy plant minikit
    si <t>RNA</t> knockdown of TRPM 7 channel abrogates Ca 2+ influx and I spont in <t>K562</t> cells. (A) detection of TRPM 7 transcripts (398 bp) by conventional RT ‐ PCR . Ten μ mol/L doxycycline ( DXC ) almost completely eliminated TRPM 7 mRNA expression. (B) immunoblots of K562 protein extracts by TRPM 7 antibody. Doxycycline concentration dependently (0.1–10 μ mol/L) reduced TRPM 7 protein expression. The results shown in A and B are representative of three independent experiments. (C) fura‐2 Ca 2+ fluorescence imaging from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing tet ‐inducible TRPM 7‐specific si RNA (si TRPM 7). n = 36 and 26, respectively. (D) density of I spont recorded by conventional whole‐cell recording from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing si TRPM 7. To facilitate the induction of TRPM 7 currents, ATP and Mg 2+ were absent in the patch pipette. The numbers of cells tested are shown in parentheses.
    Rneasy Plant Minikit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 11659 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen extraction kit
    si <t>RNA</t> knockdown of TRPM 7 channel abrogates Ca 2+ influx and I spont in <t>K562</t> cells. (A) detection of TRPM 7 transcripts (398 bp) by conventional RT ‐ PCR . Ten μ mol/L doxycycline ( DXC ) almost completely eliminated TRPM 7 mRNA expression. (B) immunoblots of K562 protein extracts by TRPM 7 antibody. Doxycycline concentration dependently (0.1–10 μ mol/L) reduced TRPM 7 protein expression. The results shown in A and B are representative of three independent experiments. (C) fura‐2 Ca 2+ fluorescence imaging from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing tet ‐inducible TRPM 7‐specific si RNA (si TRPM 7). n = 36 and 26, respectively. (D) density of I spont recorded by conventional whole‐cell recording from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing si TRPM 7. To facilitate the induction of TRPM 7 currents, ATP and Mg 2+ were absent in the patch pipette. The numbers of cells tested are shown in parentheses.
    Extraction Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 757 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen rneasy protect saliva minikit
    si <t>RNA</t> knockdown of TRPM 7 channel abrogates Ca 2+ influx and I spont in <t>K562</t> cells. (A) detection of TRPM 7 transcripts (398 bp) by conventional RT ‐ PCR . Ten μ mol/L doxycycline ( DXC ) almost completely eliminated TRPM 7 mRNA expression. (B) immunoblots of K562 protein extracts by TRPM 7 antibody. Doxycycline concentration dependently (0.1–10 μ mol/L) reduced TRPM 7 protein expression. The results shown in A and B are representative of three independent experiments. (C) fura‐2 Ca 2+ fluorescence imaging from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing tet ‐inducible TRPM 7‐specific si RNA (si TRPM 7). n = 36 and 26, respectively. (D) density of I spont recorded by conventional whole‐cell recording from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing si TRPM 7. To facilitate the induction of TRPM 7 currents, ATP and Mg 2+ were absent in the patch pipette. The numbers of cells tested are shown in parentheses.
    Rneasy Protect Saliva Minikit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen buffer rlt plus
    si <t>RNA</t> knockdown of TRPM 7 channel abrogates Ca 2+ influx and I spont in <t>K562</t> cells. (A) detection of TRPM 7 transcripts (398 bp) by conventional RT ‐ PCR . Ten μ mol/L doxycycline ( DXC ) almost completely eliminated TRPM 7 mRNA expression. (B) immunoblots of K562 protein extracts by TRPM 7 antibody. Doxycycline concentration dependently (0.1–10 μ mol/L) reduced TRPM 7 protein expression. The results shown in A and B are representative of three independent experiments. (C) fura‐2 Ca 2+ fluorescence imaging from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing tet ‐inducible TRPM 7‐specific si RNA (si TRPM 7). n = 36 and 26, respectively. (D) density of I spont recorded by conventional whole‐cell recording from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing si TRPM 7. To facilitate the induction of TRPM 7 currents, ATP and Mg 2+ were absent in the patch pipette. The numbers of cells tested are shown in parentheses.
    Buffer Rlt Plus, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 499 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen buffer rpe
    si <t>RNA</t> knockdown of TRPM 7 channel abrogates Ca 2+ influx and I spont in <t>K562</t> cells. (A) detection of TRPM 7 transcripts (398 bp) by conventional RT ‐ PCR . Ten μ mol/L doxycycline ( DXC ) almost completely eliminated TRPM 7 mRNA expression. (B) immunoblots of K562 protein extracts by TRPM 7 antibody. Doxycycline concentration dependently (0.1–10 μ mol/L) reduced TRPM 7 protein expression. The results shown in A and B are representative of three independent experiments. (C) fura‐2 Ca 2+ fluorescence imaging from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing tet ‐inducible TRPM 7‐specific si RNA (si TRPM 7). n = 36 and 26, respectively. (D) density of I spont recorded by conventional whole‐cell recording from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing si TRPM 7. To facilitate the induction of TRPM 7 currents, ATP and Mg 2+ were absent in the patch pipette. The numbers of cells tested are shown in parentheses.
    Buffer Rpe, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 164 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen lysis solution
    si <t>RNA</t> knockdown of TRPM 7 channel abrogates Ca 2+ influx and I spont in <t>K562</t> cells. (A) detection of TRPM 7 transcripts (398 bp) by conventional RT ‐ PCR . Ten μ mol/L doxycycline ( DXC ) almost completely eliminated TRPM 7 mRNA expression. (B) immunoblots of K562 protein extracts by TRPM 7 antibody. Doxycycline concentration dependently (0.1–10 μ mol/L) reduced TRPM 7 protein expression. The results shown in A and B are representative of three independent experiments. (C) fura‐2 Ca 2+ fluorescence imaging from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing tet ‐inducible TRPM 7‐specific si RNA (si TRPM 7). n = 36 and 26, respectively. (D) density of I spont recorded by conventional whole‐cell recording from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing si TRPM 7. To facilitate the induction of TRPM 7 currents, ATP and Mg 2+ were absent in the patch pipette. The numbers of cells tested are shown in parentheses.
    Lysis Solution, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 426 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    si RNA knockdown of TRPM 7 channel abrogates Ca 2+ influx and I spont in K562 cells. (A) detection of TRPM 7 transcripts (398 bp) by conventional RT ‐ PCR . Ten μ mol/L doxycycline ( DXC ) almost completely eliminated TRPM 7 mRNA expression. (B) immunoblots of K562 protein extracts by TRPM 7 antibody. Doxycycline concentration dependently (0.1–10 μ mol/L) reduced TRPM 7 protein expression. The results shown in A and B are representative of three independent experiments. (C) fura‐2 Ca 2+ fluorescence imaging from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing tet ‐inducible TRPM 7‐specific si RNA (si TRPM 7). n = 36 and 26, respectively. (D) density of I spont recorded by conventional whole‐cell recording from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing si TRPM 7. To facilitate the induction of TRPM 7 currents, ATP and Mg 2+ were absent in the patch pipette. The numbers of cells tested are shown in parentheses.

    Journal: Physiological Reports

    Article Title: TRPM7‐mediated spontaneous Ca2+ entry regulates the proliferation and differentiation of human leukemia cell line K562. TRPM7‐mediated spontaneous Ca2+ entry regulates the proliferation and differentiation of human leukemia cell line K562

    doi: 10.14814/phy2.13796

    Figure Lengend Snippet: si RNA knockdown of TRPM 7 channel abrogates Ca 2+ influx and I spont in K562 cells. (A) detection of TRPM 7 transcripts (398 bp) by conventional RT ‐ PCR . Ten μ mol/L doxycycline ( DXC ) almost completely eliminated TRPM 7 mRNA expression. (B) immunoblots of K562 protein extracts by TRPM 7 antibody. Doxycycline concentration dependently (0.1–10 μ mol/L) reduced TRPM 7 protein expression. The results shown in A and B are representative of three independent experiments. (C) fura‐2 Ca 2+ fluorescence imaging from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing tet ‐inducible TRPM 7‐specific si RNA (si TRPM 7). n = 36 and 26, respectively. (D) density of I spont recorded by conventional whole‐cell recording from untreated ([ DXC (−)] and 10 μ mol/L doxycycline‐treated [ DXC (+)] K562 cells stably expressing si TRPM 7. To facilitate the induction of TRPM 7 currents, ATP and Mg 2+ were absent in the patch pipette. The numbers of cells tested are shown in parentheses.

    Article Snippet: Reverse transcription polymerase reaction (RT‐PCR) and real‐time PCR Total RNA (about 1 μ g) was extracted from about a million of K562 cells with the total RNA extraction kit (RNeasy Minikit, Qiagen) and reverse transcribed (in a 10 μ L scale) by SuperScriptII (Thermo Fisher Scientific) with a randomized or an oligo‐dT primer to obtain the first strand DNA.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Concentration Assay, Fluorescence, Imaging, Stable Transfection, Transferring

    K562 cell growth is dependent on Ca 2+ influx mediated by TRPM 7 channel. (A) Dependency of K562 cell growth on extracellular Ca 2+ concentration evaluated at 72 h after culture. Open and filled circles indicate K562 cells untreated [ DXC (‐)] or treated with 10 μ mol/L doxycycline [ DXC (+)], respectively. (B) time courses of K562 cell growth under various conditions, i.e. DXC (−) (no doxycycline and normal Ca 2+ in RPMI medium), DXC (−)0Ca 2+ (no doxycycline and Ca 2+ free in the medium), DXC (+) (10 μ mol/L doxycycline and normal Ca 2+ in the medium) and DXC (+)0Ca 2+ (10 μ mol/L doxycycline and Ca 2+ free in the medium). In both types of experiments (A and B), K562 cells stably expressing tet ‐inducible TRPM 7‐si RNA (si TRPM 7) were used. n = 5 for each condition. * P

    Journal: Physiological Reports

    Article Title: TRPM7‐mediated spontaneous Ca2+ entry regulates the proliferation and differentiation of human leukemia cell line K562. TRPM7‐mediated spontaneous Ca2+ entry regulates the proliferation and differentiation of human leukemia cell line K562

    doi: 10.14814/phy2.13796

    Figure Lengend Snippet: K562 cell growth is dependent on Ca 2+ influx mediated by TRPM 7 channel. (A) Dependency of K562 cell growth on extracellular Ca 2+ concentration evaluated at 72 h after culture. Open and filled circles indicate K562 cells untreated [ DXC (‐)] or treated with 10 μ mol/L doxycycline [ DXC (+)], respectively. (B) time courses of K562 cell growth under various conditions, i.e. DXC (−) (no doxycycline and normal Ca 2+ in RPMI medium), DXC (−)0Ca 2+ (no doxycycline and Ca 2+ free in the medium), DXC (+) (10 μ mol/L doxycycline and normal Ca 2+ in the medium) and DXC (+)0Ca 2+ (10 μ mol/L doxycycline and Ca 2+ free in the medium). In both types of experiments (A and B), K562 cells stably expressing tet ‐inducible TRPM 7‐si RNA (si TRPM 7) were used. n = 5 for each condition. * P

    Article Snippet: Reverse transcription polymerase reaction (RT‐PCR) and real‐time PCR Total RNA (about 1 μ g) was extracted from about a million of K562 cells with the total RNA extraction kit (RNeasy Minikit, Qiagen) and reverse transcribed (in a 10 μ L scale) by SuperScriptII (Thermo Fisher Scientific) with a randomized or an oligo‐dT primer to obtain the first strand DNA.

    Techniques: Concentration Assay, Stable Transfection, Expressing

    Erythroid differentiation of K562 cell by hemin is regulated by Ca 2+ influx mediated by TRPM 7. (A) representative RT ‐ PCR experiments showing hemin‐induced γ ‐globin synthesis in K562 cells untreated (control) and treated with 10 μ mol/L FTY 720 (+ FTY ), 1 mmol/L EGTA (+ EGTA ) or 10 μ mol/L doxycycline (+dxc). In all conditions, K562 cells stably expressing tet ‐inducible TRPM 7‐si RNA (si TRPM 7) were used. The expression of γ ‐globin mRNA was greatly decreased by these procedues, while that of GAPDH stayed almost constant. (B and C) statistical evaluation of pooled data from experiments as shown in A: 10 μ mol/L FTY 720 and 1 mmol/L EGTA (B): si TRPM 7 (C). (D and E) effects of 10 μ mol/L FTY 720 ( FTY ), 1 mmol/L EGTA ( EGTA ) or 10 μ mol/L doxycycline (+dxc) on hemoglobin synthesis at rest or 3 days after hemin treatment in K562‐cells stably expressing si TRPM 7. n = 5 for each condition. * P

    Journal: Physiological Reports

    Article Title: TRPM7‐mediated spontaneous Ca2+ entry regulates the proliferation and differentiation of human leukemia cell line K562. TRPM7‐mediated spontaneous Ca2+ entry regulates the proliferation and differentiation of human leukemia cell line K562

    doi: 10.14814/phy2.13796

    Figure Lengend Snippet: Erythroid differentiation of K562 cell by hemin is regulated by Ca 2+ influx mediated by TRPM 7. (A) representative RT ‐ PCR experiments showing hemin‐induced γ ‐globin synthesis in K562 cells untreated (control) and treated with 10 μ mol/L FTY 720 (+ FTY ), 1 mmol/L EGTA (+ EGTA ) or 10 μ mol/L doxycycline (+dxc). In all conditions, K562 cells stably expressing tet ‐inducible TRPM 7‐si RNA (si TRPM 7) were used. The expression of γ ‐globin mRNA was greatly decreased by these procedues, while that of GAPDH stayed almost constant. (B and C) statistical evaluation of pooled data from experiments as shown in A: 10 μ mol/L FTY 720 and 1 mmol/L EGTA (B): si TRPM 7 (C). (D and E) effects of 10 μ mol/L FTY 720 ( FTY ), 1 mmol/L EGTA ( EGTA ) or 10 μ mol/L doxycycline (+dxc) on hemoglobin synthesis at rest or 3 days after hemin treatment in K562‐cells stably expressing si TRPM 7. n = 5 for each condition. * P

    Article Snippet: Reverse transcription polymerase reaction (RT‐PCR) and real‐time PCR Total RNA (about 1 μ g) was extracted from about a million of K562 cells with the total RNA extraction kit (RNeasy Minikit, Qiagen) and reverse transcribed (in a 10 μ L scale) by SuperScriptII (Thermo Fisher Scientific) with a randomized or an oligo‐dT primer to obtain the first strand DNA.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Stable Transfection, Expressing