Rabbit anti-NUP153 Antibody, Affinity Purified A301-788A-T from Bethyl Laboratories, Inc.

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Nature Physics
July 01 2024

Fig 1: Nup153 regulates NPC sensitivity to the mechanical directionality of the translocating protein cargo.a, Schematics of the NPC, showing the approximate location of Nup153, Nup98 and Nup214. b, Schematics of the dot blot assay. c, Dot blot membranes to assess protein binding to Nup153, Nup98 and Nup214, including a control (no Nup present). d, Dot blot intensity for each of the Nups interacting with Ig27WT, normalized by the control experiment. Each data point corresponds to the average intensity of three dots per membrane (N = 5 independent dot blot experiments). Significance levels for two-tailed t-test: NS, P > 0.05; **P < 0.01; Nup153 versus Nup214, P = 6.94 × 10−3; Nup214 versus Nup98, P = 0.76; Nup153 versus Nup98, P = 1.42 × 10−3. Boxes span from the first to the third quartile, and the horizontal line indicates the median. e, Dot blot intensity for Nup153 binding to either Ig27WT or (GS)4-Ig27WT. Significance levels fo the two-tailed paired t-test: *P < 0.05. WT versus (GS)4-WT, P = 0.04. f, Representative confocal images of U2OS cells 30 min into the recovery phase. Scale bar, 10 µm. g, Time courses of the relative nucleus-to-cytoplasm localization of R16-WT versus WT-R16 (top) and R16-Y9P versus Y9P-R16 (bottom) under normal conditions (siNT) and Nup153 knockdown (siNup153). h, Associated import rates: R16-WT (siNT) (kI = 1.35 ± 0.05 ks−1, n = 130), R16-WT (siNup153) (kI = 1.15 ± 0.05 ks−1, n = 108); WT-R16 (siNT) (kI = 1.20 ± 0.05 ks−1, n = 179), WT-R16 (siNup153) (kI = 1.09 ± 0.05 ks−1, n = 98); R16-Y9P (siNT) (kI = 1.51 ± 0.06 ks−1, n = 92), R16-Y9P (siNup153) (kI = 0.93 ± 0.06 ks−1, n = 57); Y9P-R16 (siNT) (kI = 1.07 ± 0.04 ks−1, n = 100), Y9P-R16 (siNup153) (kI = 0.83 ± 0.09 ks−1, n = 33). Significance levels for the two-tailed Mann–Whitney non-parametric test: NS, P > 0.05; *P < 0.05; **P < 0.01; ****P < 0.0001; siNT R16-WT versus siNup153 R16-WT, P = 2.14 × 10−3; siNT WT-R16 versus siNup153 WT-R16, P = 0.02; siNT R16-WT versus siNT WT-R16, P = 0.03; siNup153 R16-WT versus siNUP153 WT-R16, P = 0.25. siNT R16-Y9P versus siNup153 R16-Y9P, P = 5.70 × 10−9; siNT Y9P-R16 versus siNup153 Y9P-R16, P = 2.50 × 10−3; siNT R16-Y9P versus siNT Y9P-R16, P = 1.26 × 10−7; siNup153 R16-Y9P versus siNUP153 Y9P-R16, P = 0.17. i, Representative confocal image of U2OS cells. Scale bar, 10 µm. j, Time courses of the relative nucleus-to-cytoplasm localization of WT and (GS)4-WT under normal conditions (siNT) and Nup153 knockdown (siNup153). k, Associated import rates; WT (siNT) (kI = 1.61 ± 0.05 ks−1, n = 147), WT (siNup153) (kI = 1.22 ± 0.05 ks−1, n = 118); (GS)4-WT (siNT) (kI = 2.02 ± 0.05 ks−1, n = 169), (GS)4-WT (siNup153) (kI = 1.28 ± 0.04 ks−1, n = 122). Significance levels for the two-tailed Mann–Whitney non-parametric test: NS, P > 0.05; ****P < 0.0001; siNT WT versus siNup153 WT, P = 7.74 × 10−7; siNT (GS)4-WT versus siNUP153 (GS)4-WT, P = 2.50 × 10−28; siNT WT versus siNT (GS)4-WT, P = 1.42 × 10−6; siNup153 WT versus siNup153 (GS)4-WT, P = 0.59. l, Directionality calculated as the ratio between the average import rate (top) and accumulation (bottom) for translocating R16-Ig27Y9P versus Ig27Y9P-R16 constructs, measured when silencing individual FG-Nups. Error bars indicate s.e.m. m, Schematic of the proposed role of Nup153 in identifying locally unstructured protein regions. Significance levels for two-tailed t-test (d), two-tailed paired t-test (e), Mann–Whitney non-parametric test (h,k): NS, P > 0.05; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. All points and bar plots indicate mean ± s.e.m.
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Nature Physics
July 01 2024

Fig 2: Knockdown of Nup153 decreases nuclear import kinetics but does not affect NPC mechanosensitivity.(A) Representative confocal images of U2OS cells 30 min in the recovery phase, scale bar 10 µm. (B) Average time courses of the relative nucleus-to-cytoplasm localization of V13P and WT under normal conditions (siNT) and Nup153 knockdown (siNup153). (C) Import rates calculated from fits to the recovery time courses. V13P (siNT) (kI = 2.32 ± 0.09 ks−1, n = 82), V13P (siNup153) (kI = 1.35 ± 0.04 ks−1, n = 100); WT (siNT) (kI = 1.61 ± 0.05 ks−1, n = 147), WT (siNup153) (kI = 1.22 ± 0.05 ks−1, n = 118). Significance levels for two-tailed Mann-Whitney non-parametric test NS > 0.05, *** P ≤ 0.001, **** P ≤ 0.0001. siNT V13P vs. siNup153 V13P, P = 3.21 × 10−29; siNT WT vs. siNup153 WT, P = 9.80 × 10−10; siNT V13P vs. siNT WT, P = 3.14 × 10−11. siNup153 V13P vs. siNup153 WT, P = 6.78 × 10−5. All points and bar plots indicate mean ± SEM.
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Nature Physics
April 30 2026

Fig 3: Downregulation of Nup153 reduces the sensitivity of the NPC to the local mechanical asymmetry of imported protein cargos.(A-C) Nuclear import kinetics of the mCherry protein when imported from the N- or C-terminus upon downregulating Nup153. (A) Representative confocal images of U2OS cells after 20 min of the recovery phase, scale bar 10 µm. (B) Average time-courses of the nucleus-to-cytoplasm localisation of the “Import forward” and “Import reverse” mCherry in the control group (siNT) and knocking down Nup153 (siNup153). (C) Relative nuclear accumulation (left) and import rate calculated from fits to the recovery time courses. Bars indicate mean. Accumulation: N-term (siNT) Ke = 2.21 ± 0.07; N-term (siNup153) Ke = 1.00 ± 0.04; C-term (siNT) Ke = 1.14 ± 0.03; C-term (siNup153) Ke = 0.83 ± 0.02. Import rate: N-term (siNT) kI = 3.95 ± 0.14 ks−1; N-term (siNup153) kI = 1.59 ± 0.08 ks−1; C-term (siNT) kI = 0.83 ± 0.02 ks−1; C-term (siNup153) kI = 1.13 ± 0.04 ks−1. Significance levels for two-tailed Mann-Whitney non-parametric test. Accumulation: N-term (siNT) vs. C-term (siNT) P = 0; N-term (siNup153) vs. C-term (siNup153) P = P = 4.00x10−4; N-term (siNT) vs. N-term (siNup153) P = 0; C-term (siNT) vs. C-term (siNup153) P = 9.9x10−12. Import rate: N-term (siNT) vs. C-term (siNT) P = 0; N-term (siNup153) vs. C-term (siNup153) P = 3.0x10−6; N-term (siNT) vs. N-term (siNup153) P = 0; N-term (siNup153) vs. C-term (siNup153) P = 6.7x10−16. n = 103 (siNT N-term); n = 100 (siNT C-term); n = 85 (siNUP153 N-term); n = 91 (siNUP153 C-term). (D-F) Nuclear import kinetics of the Ig27 when imported from the N- or C- terminus upon downregulating Nup153. (D) Representative confocal images of U2OS cells after 20 min of the recovery phase, scale bar 10 µm. (E) Average time-courses of the nucleus-to-cytoplasm localisation of the “Import forward” and “Import reverse” Ig27 in the control group (siNT) and knocking down Nup153 (siNup153). (F) Relative nuclear accumulation (left) and import rate calculated from fits to the recovery time courses. Bars indicate mean. Accumulation: N-term (siNT) Ke = 1.43 ± 0.04; C-term (siNT) Ke = 1.15 ± 0.03; N-term (siNUP153) Ke = 0.64 ± 0.02; C-term (siNup153) Ke = 0.64 ± 0.02. Import rate: N-term (siNT) kI = 1.63 ± 0.06 ks−1; C-term (siNT) kI = 0.97 ± 0.03 ks−1; N-term (siNUP153) kI = 0.60 ± 0.02 ks−1; C-term (siNup153) kI = 0.44 ± 0.02 ks−1. Significance levels for two-tailed Mann-Whitney non-parametric test. Accumulation: N-term (siNT) vs. C-term (siNT) P = 5.2x10−9; N-term (siNup153) vs. C-term (siNup153) P = 0.71; N-term (siNT) vs. N-term (siNup153) P = 0; C-term (siNT) vs. C-term (siNup153) P = 0. Import rate: N-term (siNT) vs. C-term (siNT) P = 1.3x10−13; N-term (siNup153) vs. C-term (siNup153) P = 5.2x10−8; N-term (siNT) vs. N-term (siNup153) P = 0; C-term (siNup153) vs. C-term (siNT) P = 0. n = 103 (siNT N-term); n = 101 (siNT C-term); n = 85 (siNUP153 N-term); n = 91 (siNUP153 C-term). (G-I) Nuclear import kinetics of the Spy0128 E117A when imported from the N- or C-terminus upon knocking down Nup153. (G) Representative confocal images of U2OS cells after 20 min of the recovery phase, scale bar 10 µm. (H) Average time-courses of the nucleus-to-cytoplasm localisation of the “Import forward” and “Import reverse” Spy0128 E117A in the control group (siNT) and knocking down Nup153 (siNup153). (I) Relative nuclear accumulation (left) and import rate calculated from fits to the recovery time courses. Bars indicate mean. Accumulation: N-term (siNT) Ke = 0.66 ± 0.22; C-term (siNT) Ke = 0.54 ± 0.02; N-term (siNUP153) Ke = 0.40 ± 0.03; C-term (siNup153) Ke = 0.41 ± 0.06. Import rate: N-term (siNT) kI = 0.63 ± 0.03 ks−1; C-term (siNT) kI = 0.29 ± 0.02 ks−1; N-term (siNUP153) kI = 0.34 ± 0.02 ks−1; C-term (siNup153) kI = 0.16 ± 0.02 ks−1. Significance levels for two-tailed Mann-Whitney non-parametric test. Accumulation: N-term (siNT) vs. C-term (siNT) P = 1.0x10−3; N-term (siNup153) vs. C-term (siNup153) P = 0.40; N-term (siNT) vs. N-term (siNup153) P = 6.9x10−10; C-term (siNT) vs. C-term (siNup153) P = 4.8x10−3. Import rate: N-term (siNT) vs. C-term (siNT) P = 1.2x10−13; N-term (siNup153) vs. C-term (siNup153) P = 4.9x10−5; N-term (siNT) vs. N-term (siNup153) P = 3.2x10−11; C-term (siNup153) vs. C-term (siNT) P = 1.0x10−3. n = 81 (siNT N-term); n = 37 (siNT C-term); n = 37 (siNUP153 N-term); n = 10 (siNUP153 C-term). (J) Percentage of directionality in nuclear accumulation (calculated as the ratio between the N-term and C-term accumulation) for the siNT control group and siNup153. Error bars indicate SEM. (K) Schematics of the proposed role of NUP153 in sensing mechanically labile termini to accelerate nuclear transport. Source data

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Rabbit anti-NUP153 Antibody, Affinity Purified

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Bethyl Laboratories, Inc.'s Rabbit anti-NUP153 Antibody, Affinity Purified is a Rabbit Polyclonal antibody. This antibody has been shown to work in applications such as: Precipitation, Immunocytochemistry, Immunofluorescence, Immunoprecipitation, and Western Blot. The Rabbit anti-NUP153 Antibody, Affinity Purified was generated using NUP153 as the antigen and it reacts with Homo Sapiens, and Human.

Description

The epitope recognized by A301-788A-T maps to a region between residue 50 and 100 of human nucleoporin 153kDa using the numbering given in entry NP_005115.2 (GeneID 9972)

Nature Physics

Nature Physics

Nature Physics

Rabbit anti-NUP153 Antibody, Affinity Purified from Bethyl Laboratories, Inc.

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Bethyl Laboratories, Inc.

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Rabbit anti-NUP153 Antibody, Affinity Purified from Bethyl Laboratories, Inc.

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