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Use the natural cubic spline convention appropriate to the interior design knots: the spline is linear on $[a,x_1]$ and $[x_n,b]$. We also need $n\ge 2$. These points will be important for uniqueness.

A cubic spline is a function in $C^2[a,b]$ whose restriction to each of $[a,x_1]$, $[x_1,x_2]$, through $[x_n,b]$ is a polynomial of degree at most three. A spline knot is a junction between consecutive pieces. For a natural cubic spline, the two exterior pieces are affine functions; equivalently, one uses the natural endpoint conditions $g^{\prime \prime }(x_1)=g^{\prime \prime }(x_n)=0$ on the spline over $[x_1,x_n]$ and then extends linearly with matching derivatives. Merely prescribing $g^{\prime \prime }(a)=g^{\prime \prime }(b)=0$ on otherwise arbitrary exterior cubic pieces is a different condition and does not provide the unique interpolant invoked in this question.

The natural cubic spline interpolant $N\mathbf{v}$ to values $\mathbf{v}=(v_1,\dots ,v_n{)}^T$ is the natural cubic spline satisfying $(N\mathbf{v})(x_i)=v_i$. Its existence and uniqueness for $n\ge 2$ are the interpolant fact allowed in the PDF. The interpolation map $N$ is a linear map: a linear combination of interpolants satisfies the same spline, boundary and value conditions, so uniqueness identifies it with the interpolant to the corresponding linear combination of value vectors.

Let $g=N\mathbf{v}$, let $\tilde{g}\in C^2[a,b]$ have the same values, and put $r=\tilde{g}-g$. Then $r(x_i)=0$. We prove the orthogonalityApply integration by parts twice on each polynomial interval. On an interior interval, $g^{\prime \prime \prime \prime }=0$, andThe $g^{\prime \prime \prime }r$ terms vanish because $r$ vanishes at the knots. The $g^{\prime \prime }{r}^{\prime }$ terms cancel at internal knots because $g^{\prime \prime }$ and $r^{\prime }$ are continuous, and vanish at $x_1,x_n$ because $g^{\prime \prime }$ is zero there. On the exterior intervals, $g^{\prime \prime }=0$ identically. Summing therefore gives the claimed orthogonality without assuming that $g^{\prime \prime \prime }$ is continuous across knots.

Expanding the square now gives the minimum roughness property of the natural cubic spline interpolantEquality forces $r^{\prime \prime }=0$ everywhere, since $r^{\prime \prime }$ is continuous. Thus $r$ is an affine function. It vanishes at two distinct design points, so $r=0$, proving equality if and only if $\tilde{g}=g$.

For the penalized fit, take the allowed spline roughness penalty matrix $\Gamma$ to be a symmetric matrix with ${\mathbf{z}}^T\Gamma \mathbf{z}\ge 0$; thus it is a positive semidefinite matrix. If its supplied representative were not symmetric, its symmetric part defines the same quadratic form and is all that is needed. For any $\tilde{g}$, let $\mathbf{v}=(\tilde{g}(x_i){)}_{i=1}^n$. Replacing it by $N\mathbf{v}$ preserves every fitted value and can only decrease the second derivative roughness penalty. Therefore it is enough to minimizeover ${\mathbb{R}}^n$. The matrix $A_{\lambda }=I_n+\lambda \Gamma$ is a positive-definite matrix, sinceCompleting the quadratic or differentiating gives the unique solutionThis is the cubic smoothing spline. More explicitly, the normal equations are $(I_n+\lambda \Gamma ){\hat{\mathbf{v}}}_{\lambda }=\mathbf{Y}$, andFor an arbitrary function with fitted vector $\mathbf{v}$, the earlier orthogonality also givesBoth terms are nonnegative. Equality forces $\mathbf{v}={\hat{\mathbf{v}}}_{\lambda }$ and, by the proved interpolation equality case, $\tilde{g}=N\mathbf{v}={\hat{g}}_{\lambda }$. This proves both existence in $C^2[a,b]$ and uniqueness over the entire stated function class, not just over splines.

The argument is deterministic for each observed $\mathbf{Y}$. The fixed-design nonparametric regression and homoscedasticity assumptions motivate the squared-error loss, but Gaussian errors are not required.

At least two distinct design points are required for uniqueness. Question 4 does not explicitly restate $n\ge 2$, and this hypothesis is necessary. With one design point, every function $\tilde{g}(x)=Y_1+c(x-x_1)$ has zero residual and zero second derivative roughness penalty, for any real $c$. Thus the printed unrestricted uniqueness conclusion would be false for $n=1$. The corrected theorem assumes at least two distinct knots and the stated linear-tail convention. At $\lambda =0$, uniqueness over all of $C^2[a,b]$ also fails; the stipulated $\lambda >0$ eliminates that degeneracy.

The nonnegative functional $J(g)={\int }_a^b(g^{\prime \prime }(x){)}^{2}dx$. Its null space on $C^2[a,b]$ consists of affine functions. The natural cubic spline interpolant uniquely minimizes this penalty among functions matching at least two prescribed knot values. It is a squared second derivative penalty, not the exact geometric curvature energy of a graph.