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primary dermal fibroblast normal  (ATCC)


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    ATCC primary dermal fibroblast normal
    CCDC134 deficiency selectively impaired TLR-mediated immune responses. (A) Immunoprecipitates (IP) and whole-cell extracts (WCE) from HEK293T cells transfected as indicated. SH: Strep-HA tag. (B and C) Immunoblots of indicated knockout THP1 cells unstimulated (B) or stimulated with R848 (5 µg/ml, for 0–1 h) (C). (D) Indicated knockout THP1 DUAL reporter cells stimulated with R848 (5 μg/ml) or LPS (0.1 μg/ml) for 24 h. Supernatants were analyzed for ISRE and NF-κB reporter activity. Untr.: untreated. (E) Immunoblots of cell lysates from indicated knockout THP1 cells treated with EndoH (H) or PNGase F (F). (F) TNFα production of indicated knockout THP1 cells stimulated for 24 h with LPS (0.1 μg/ml), R848 (5 μg/ml), Pam3CSK4 (0.1 μg/ml) or Pam2CSK4 (0.01 μg/ml). Untr.: untreated. (G) Immunoblots of cell lysates from indicated knockout U937 cells. (H) IL-6 (left panel) and TNFα (right panel) production of indicated knockout U937 cells differentiated with 200 nM of PMA for 24 h before stimulation with LPS (0.1 μg/ml), Pam3CSK4 (Pam3.) (1 μg/ml), or Pam2CSK4 (Pam2.) (0.1 μg/ml) for 24 h. Untr.: untreated. (I and J) Immunoblots of cell lysates from indicated knockout U937 cells differentiated with 200 nM of PMA for 48 h, treated with EndoH (H) or PNGase F (F) (I) or from indicated knockout human primary dermal <t>fibroblast</t> cells (J). Mat.: Mature form; Imat.: Immature form. (K) IL-6 production of indicated knockout human primary dermal fibroblast cells stimulated with LPS (0.1 μg/ml) or Pam2CSK4 (Pam2.) (0.01 μg/ml) for 24 h. Untr.: untreated. In A–C, E, G, I, and J, data are representative of two independent experiments. In D, F, H, and K, data show mean ± SD of three stimulation replicates from one experiment representative of three independent experiments. Source data are available for this figure: .
    Primary Dermal Fibroblast Normal, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "CCDC134 controls TLR biogenesis through the ER chaperone Gp96"

    Article Title: CCDC134 controls TLR biogenesis through the ER chaperone Gp96

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20240825

    CCDC134 deficiency selectively impaired TLR-mediated immune responses. (A) Immunoprecipitates (IP) and whole-cell extracts (WCE) from HEK293T cells transfected as indicated. SH: Strep-HA tag. (B and C) Immunoblots of indicated knockout THP1 cells unstimulated (B) or stimulated with R848 (5 µg/ml, for 0–1 h) (C). (D) Indicated knockout THP1 DUAL reporter cells stimulated with R848 (5 μg/ml) or LPS (0.1 μg/ml) for 24 h. Supernatants were analyzed for ISRE and NF-κB reporter activity. Untr.: untreated. (E) Immunoblots of cell lysates from indicated knockout THP1 cells treated with EndoH (H) or PNGase F (F). (F) TNFα production of indicated knockout THP1 cells stimulated for 24 h with LPS (0.1 μg/ml), R848 (5 μg/ml), Pam3CSK4 (0.1 μg/ml) or Pam2CSK4 (0.01 μg/ml). Untr.: untreated. (G) Immunoblots of cell lysates from indicated knockout U937 cells. (H) IL-6 (left panel) and TNFα (right panel) production of indicated knockout U937 cells differentiated with 200 nM of PMA for 24 h before stimulation with LPS (0.1 μg/ml), Pam3CSK4 (Pam3.) (1 μg/ml), or Pam2CSK4 (Pam2.) (0.1 μg/ml) for 24 h. Untr.: untreated. (I and J) Immunoblots of cell lysates from indicated knockout U937 cells differentiated with 200 nM of PMA for 48 h, treated with EndoH (H) or PNGase F (F) (I) or from indicated knockout human primary dermal fibroblast cells (J). Mat.: Mature form; Imat.: Immature form. (K) IL-6 production of indicated knockout human primary dermal fibroblast cells stimulated with LPS (0.1 μg/ml) or Pam2CSK4 (Pam2.) (0.01 μg/ml) for 24 h. Untr.: untreated. In A–C, E, G, I, and J, data are representative of two independent experiments. In D, F, H, and K, data show mean ± SD of three stimulation replicates from one experiment representative of three independent experiments. Source data are available for this figure: .
    Figure Legend Snippet: CCDC134 deficiency selectively impaired TLR-mediated immune responses. (A) Immunoprecipitates (IP) and whole-cell extracts (WCE) from HEK293T cells transfected as indicated. SH: Strep-HA tag. (B and C) Immunoblots of indicated knockout THP1 cells unstimulated (B) or stimulated with R848 (5 µg/ml, for 0–1 h) (C). (D) Indicated knockout THP1 DUAL reporter cells stimulated with R848 (5 μg/ml) or LPS (0.1 μg/ml) for 24 h. Supernatants were analyzed for ISRE and NF-κB reporter activity. Untr.: untreated. (E) Immunoblots of cell lysates from indicated knockout THP1 cells treated with EndoH (H) or PNGase F (F). (F) TNFα production of indicated knockout THP1 cells stimulated for 24 h with LPS (0.1 μg/ml), R848 (5 μg/ml), Pam3CSK4 (0.1 μg/ml) or Pam2CSK4 (0.01 μg/ml). Untr.: untreated. (G) Immunoblots of cell lysates from indicated knockout U937 cells. (H) IL-6 (left panel) and TNFα (right panel) production of indicated knockout U937 cells differentiated with 200 nM of PMA for 24 h before stimulation with LPS (0.1 μg/ml), Pam3CSK4 (Pam3.) (1 μg/ml), or Pam2CSK4 (Pam2.) (0.1 μg/ml) for 24 h. Untr.: untreated. (I and J) Immunoblots of cell lysates from indicated knockout U937 cells differentiated with 200 nM of PMA for 48 h, treated with EndoH (H) or PNGase F (F) (I) or from indicated knockout human primary dermal fibroblast cells (J). Mat.: Mature form; Imat.: Immature form. (K) IL-6 production of indicated knockout human primary dermal fibroblast cells stimulated with LPS (0.1 μg/ml) or Pam2CSK4 (Pam2.) (0.01 μg/ml) for 24 h. Untr.: untreated. In A–C, E, G, I, and J, data are representative of two independent experiments. In D, F, H, and K, data show mean ± SD of three stimulation replicates from one experiment representative of three independent experiments. Source data are available for this figure: .

    Techniques Used: Transfection, Western Blot, Knock-Out, Activity Assay



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    CCDC134 deficiency selectively impaired TLR-mediated immune responses. (A) Immunoprecipitates (IP) and whole-cell extracts (WCE) from HEK293T cells transfected as indicated. SH: Strep-HA tag. (B and C) Immunoblots of indicated knockout THP1 cells unstimulated (B) or stimulated with R848 (5 µg/ml, for 0–1 h) (C). (D) Indicated knockout THP1 DUAL reporter cells stimulated with R848 (5 μg/ml) or LPS (0.1 μg/ml) for 24 h. Supernatants were analyzed for ISRE and NF-κB reporter activity. Untr.: untreated. (E) Immunoblots of cell lysates from indicated knockout THP1 cells treated with EndoH (H) or PNGase F (F). (F) TNFα production of indicated knockout THP1 cells stimulated for 24 h with LPS (0.1 μg/ml), R848 (5 μg/ml), Pam3CSK4 (0.1 μg/ml) or Pam2CSK4 (0.01 μg/ml). Untr.: untreated. (G) Immunoblots of cell lysates from indicated knockout U937 cells. (H) IL-6 (left panel) and TNFα (right panel) production of indicated knockout U937 cells differentiated with 200 nM of PMA for 24 h before stimulation with LPS (0.1 μg/ml), Pam3CSK4 (Pam3.) (1 μg/ml), or Pam2CSK4 (Pam2.) (0.1 μg/ml) for 24 h. Untr.: untreated. (I and J) Immunoblots of cell lysates from indicated knockout U937 cells differentiated with 200 nM of PMA for 48 h, treated with EndoH (H) or PNGase F (F) (I) or from indicated knockout human primary dermal <t>fibroblast</t> cells (J). Mat.: Mature form; Imat.: Immature form. (K) IL-6 production of indicated knockout human primary dermal fibroblast cells stimulated with LPS (0.1 μg/ml) or Pam2CSK4 (Pam2.) (0.01 μg/ml) for 24 h. Untr.: untreated. In A–C, E, G, I, and J, data are representative of two independent experiments. In D, F, H, and K, data show mean ± SD of three stimulation replicates from one experiment representative of three independent experiments. Source data are available for this figure: .
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    CCDC134 deficiency selectively impaired TLR-mediated immune responses. (A) Immunoprecipitates (IP) and whole-cell extracts (WCE) from HEK293T cells transfected as indicated. SH: Strep-HA tag. (B and C) Immunoblots of indicated knockout THP1 cells unstimulated (B) or stimulated with R848 (5 µg/ml, for 0–1 h) (C). (D) Indicated knockout THP1 DUAL reporter cells stimulated with R848 (5 μg/ml) or LPS (0.1 μg/ml) for 24 h. Supernatants were analyzed for ISRE and NF-κB reporter activity. Untr.: untreated. (E) Immunoblots of cell lysates from indicated knockout THP1 cells treated with EndoH (H) or PNGase F (F). (F) TNFα production of indicated knockout THP1 cells stimulated for 24 h with LPS (0.1 μg/ml), R848 (5 μg/ml), Pam3CSK4 (0.1 μg/ml) or Pam2CSK4 (0.01 μg/ml). Untr.: untreated. (G) Immunoblots of cell lysates from indicated knockout U937 cells. (H) IL-6 (left panel) and TNFα (right panel) production of indicated knockout U937 cells differentiated with 200 nM of PMA for 24 h before stimulation with LPS (0.1 μg/ml), Pam3CSK4 (Pam3.) (1 μg/ml), or Pam2CSK4 (Pam2.) (0.1 μg/ml) for 24 h. Untr.: untreated. (I and J) Immunoblots of cell lysates from indicated knockout U937 cells differentiated with 200 nM of PMA for 48 h, treated with EndoH (H) or PNGase F (F) (I) or from indicated knockout human primary dermal <t>fibroblast</t> cells (J). Mat.: Mature form; Imat.: Immature form. (K) IL-6 production of indicated knockout human primary dermal fibroblast cells stimulated with LPS (0.1 μg/ml) or Pam2CSK4 (Pam2.) (0.01 μg/ml) for 24 h. Untr.: untreated. In A–C, E, G, I, and J, data are representative of two independent experiments. In D, F, H, and K, data show mean ± SD of three stimulation replicates from one experiment representative of three independent experiments. Source data are available for this figure: .
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    CCDC134 deficiency selectively impaired TLR-mediated immune responses. (A) Immunoprecipitates (IP) and whole-cell extracts (WCE) from HEK293T cells transfected as indicated. SH: Strep-HA tag. (B and C) Immunoblots of indicated knockout THP1 cells unstimulated (B) or stimulated with R848 (5 µg/ml, for 0–1 h) (C). (D) Indicated knockout THP1 DUAL reporter cells stimulated with R848 (5 μg/ml) or LPS (0.1 μg/ml) for 24 h. Supernatants were analyzed for ISRE and NF-κB reporter activity. Untr.: untreated. (E) Immunoblots of cell lysates from indicated knockout THP1 cells treated with EndoH (H) or PNGase F (F). (F) TNFα production of indicated knockout THP1 cells stimulated for 24 h with LPS (0.1 μg/ml), R848 (5 μg/ml), Pam3CSK4 (0.1 μg/ml) or Pam2CSK4 (0.01 μg/ml). Untr.: untreated. (G) Immunoblots of cell lysates from indicated knockout U937 cells. (H) IL-6 (left panel) and TNFα (right panel) production of indicated knockout U937 cells differentiated with 200 nM of PMA for 24 h before stimulation with LPS (0.1 μg/ml), Pam3CSK4 (Pam3.) (1 μg/ml), or Pam2CSK4 (Pam2.) (0.1 μg/ml) for 24 h. Untr.: untreated. (I and J) Immunoblots of cell lysates from indicated knockout U937 cells differentiated with 200 nM of PMA for 48 h, treated with EndoH (H) or PNGase F (F) (I) or from indicated knockout human primary dermal <t>fibroblast</t> cells (J). Mat.: Mature form; Imat.: Immature form. (K) IL-6 production of indicated knockout human primary dermal fibroblast cells stimulated with LPS (0.1 μg/ml) or Pam2CSK4 (Pam2.) (0.01 μg/ml) for 24 h. Untr.: untreated. In A–C, E, G, I, and J, data are representative of two independent experiments. In D, F, H, and K, data show mean ± SD of three stimulation replicates from one experiment representative of three independent experiments. Source data are available for this figure: .
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    CCDC134 deficiency selectively impaired TLR-mediated immune responses. (A) Immunoprecipitates (IP) and whole-cell extracts (WCE) from HEK293T cells transfected as indicated. SH: Strep-HA tag. (B and C) Immunoblots of indicated knockout THP1 cells unstimulated (B) or stimulated with R848 (5 µg/ml, for 0–1 h) (C). (D) Indicated knockout THP1 DUAL reporter cells stimulated with R848 (5 μg/ml) or LPS (0.1 μg/ml) for 24 h. Supernatants were analyzed for ISRE and NF-κB reporter activity. Untr.: untreated. (E) Immunoblots of cell lysates from indicated knockout THP1 cells treated with EndoH (H) or PNGase F (F). (F) TNFα production of indicated knockout THP1 cells stimulated for 24 h with LPS (0.1 μg/ml), R848 (5 μg/ml), Pam3CSK4 (0.1 μg/ml) or Pam2CSK4 (0.01 μg/ml). Untr.: untreated. (G) Immunoblots of cell lysates from indicated knockout U937 cells. (H) IL-6 (left panel) and TNFα (right panel) production of indicated knockout U937 cells differentiated with 200 nM of PMA for 24 h before stimulation with LPS (0.1 μg/ml), Pam3CSK4 (Pam3.) (1 μg/ml), or Pam2CSK4 (Pam2.) (0.1 μg/ml) for 24 h. Untr.: untreated. (I and J) Immunoblots of cell lysates from indicated knockout U937 cells differentiated with 200 nM of PMA for 48 h, treated with EndoH (H) or PNGase F (F) (I) or from indicated knockout human primary dermal fibroblast cells (J). Mat.: Mature form; Imat.: Immature form. (K) IL-6 production of indicated knockout human primary dermal fibroblast cells stimulated with LPS (0.1 μg/ml) or Pam2CSK4 (Pam2.) (0.01 μg/ml) for 24 h. Untr.: untreated. In A–C, E, G, I, and J, data are representative of two independent experiments. In D, F, H, and K, data show mean ± SD of three stimulation replicates from one experiment representative of three independent experiments. Source data are available for this figure: .

    Journal: The Journal of Experimental Medicine

    Article Title: CCDC134 controls TLR biogenesis through the ER chaperone Gp96

    doi: 10.1084/jem.20240825

    Figure Lengend Snippet: CCDC134 deficiency selectively impaired TLR-mediated immune responses. (A) Immunoprecipitates (IP) and whole-cell extracts (WCE) from HEK293T cells transfected as indicated. SH: Strep-HA tag. (B and C) Immunoblots of indicated knockout THP1 cells unstimulated (B) or stimulated with R848 (5 µg/ml, for 0–1 h) (C). (D) Indicated knockout THP1 DUAL reporter cells stimulated with R848 (5 μg/ml) or LPS (0.1 μg/ml) for 24 h. Supernatants were analyzed for ISRE and NF-κB reporter activity. Untr.: untreated. (E) Immunoblots of cell lysates from indicated knockout THP1 cells treated with EndoH (H) or PNGase F (F). (F) TNFα production of indicated knockout THP1 cells stimulated for 24 h with LPS (0.1 μg/ml), R848 (5 μg/ml), Pam3CSK4 (0.1 μg/ml) or Pam2CSK4 (0.01 μg/ml). Untr.: untreated. (G) Immunoblots of cell lysates from indicated knockout U937 cells. (H) IL-6 (left panel) and TNFα (right panel) production of indicated knockout U937 cells differentiated with 200 nM of PMA for 24 h before stimulation with LPS (0.1 μg/ml), Pam3CSK4 (Pam3.) (1 μg/ml), or Pam2CSK4 (Pam2.) (0.1 μg/ml) for 24 h. Untr.: untreated. (I and J) Immunoblots of cell lysates from indicated knockout U937 cells differentiated with 200 nM of PMA for 48 h, treated with EndoH (H) or PNGase F (F) (I) or from indicated knockout human primary dermal fibroblast cells (J). Mat.: Mature form; Imat.: Immature form. (K) IL-6 production of indicated knockout human primary dermal fibroblast cells stimulated with LPS (0.1 μg/ml) or Pam2CSK4 (Pam2.) (0.01 μg/ml) for 24 h. Untr.: untreated. In A–C, E, G, I, and J, data are representative of two independent experiments. In D, F, H, and K, data show mean ± SD of three stimulation replicates from one experiment representative of three independent experiments. Source data are available for this figure: .

    Article Snippet: HEK293T cells (cat. CRL-3216), THP1 cells (cat. TIB-202), Raw 264.7 (cat. TIB-71) and Primary Dermal Fibroblast Normal; Human, Neonatal (HDFn) (cat. PCS-201-010) were purchased from ATCC.

    Techniques: Transfection, Western Blot, Knock-Out, Activity Assay